# Patent

## Title
Protective proteins of S. agalactiae, combinations thereof and methods of using the same

## Abstract

The invention relates to a composition comprising at least two protective proteins against Streptococcus agalactiae (S. agalactiae) or functionally active variant thereof; a protective peptide against S. agalactiae; one or more nucleic acid(s) encoding the at least two proteins and/or the protective peptide; a method of producing the composition; a pharmaceutical composition, especially a vaccine, comprising the composition and/or at least one protective peptide; methods for producing antibodies; a mixture of antibodies against the at least two proteins of the composition; the use of the composition and/or at least one protective peptide and/or one or more nucleic acid(s) for the manufacture of a medicament for the immunization or treatment of a subject; methods of diagnosing a S. agalactiae infection; a method for identifying a ligand capable of binding the composition and/or at least one protective peptide; and the use of the composition and/or at least one protective peptide for the isolation and/or purification and/or identification of an interaction partner of the composition and/or peptide.

## Background
CROSS REFERENCE TO RELATED APPLICATIONS
This application is a divisional of U.S. application Ser. No. 12/522,636, filed Nov. 18, 2009, now U.S. Pat. No. 8,343,510 which is the U.S. National Stage of International Application No. PCT/EP2008/050227, filed Jan. 10, 2008, which claims the benefit of European Patent Application No. 07000602.8, filed Jan. 12, 2007, each of which is hereby incorporated by reference.


The invention relates to a composition comprising at least two protective proteins against Streptococcus agalactiae (S. agalactiae) or functionally active variants thereof; a protective peptide against S. agalactiae; one or more nucleic acid(s) encoding the at least two proteins and/or the protective peptide; a method of producing the composition; a pharmaceutical composition, especially a vaccine, comprising the composition and/or at least one protective peptide; methods for producing antibodies; a mixture of antibodies against the at least two proteins of the composition; the use of the composition and/or at least one protective peptide and/or one or more nucleic acid(s) for the manufacture of a medicament for the immunization or treatment of a subject; methods of diagnosing a S. agalactiae infection; a method for identifying a ligand capable of binding the composition and/or at least one protective peptide; and the use of the composition and/or at least one protective peptide for the isolation and/or purification and/or identification of an interaction partner of the composition and/or peptide.
S. agalactiae is an encapsulated gram-positive bacterium, which belongs to the Group B Streptococci (GBS) based on its haemolysis pattern on blood agar. Capsules form the basis for classifying GBS into nine distinct serotypes. Most of them have been shown to cause serious diseases, and the two most common serotypes—type III and V—are estimated to account for the majority ( ̃80%) of invasive diseases worldwide. The ranking and serotype prevalence differs by age group and geographic area.
Streptococcus agalactiae is a frequent cause of infections in neonates, pregnant women and in chronically ill and elderly patients. In newborns Group B Streptococcus even represents the predominant pathogen in the United States causing life threatening diseases, such as sepsis, pneumonia and meningitis. GBS diseases are associated with a high mortality rate ( ̃5%) and a large percentage ( ̃20%) of children surviving GBS infections becomes permanently handicapped with hearing, learning and visual disabilities.
Newborns usually acquire the pathogen during delivery from their GBS-colonized mothers. Twenty-five to 40% of pregnant women are colonized with GBS, but are asymptomatic. Due to vertical transmission during birth, 50-70% of neonates born to colonized women—that is approximately 10-25% of all newborns—become colonized by GBS during delivery which is a prerequisite for infection and disease. In the United States, GBS infections affect 1-5 newborns/1,000 live births. Pre-term infants are at the highest risk for invasive disease due to their immature immune system and the low level of maternal antibody transfer before the 34th pregnancy week.
GBS disease occurs throughout the world. The highest prevalence of invasive disease in newborns occurs in Western countries, due to the elimination and reduction of other infectious agents and also due to the increased survival of very immature newborns. Before prevention by intrapartum antibiotic treatment was introduced, about 17,000 cases of invasive GBS diseases (sepsis, pneumonia and/or meningitis) were reported in the US annually. The rates of serious GBS infections are higher among newborns than among any other age group. Nonetheless, serious Group B streptococcal infections occur in other age groups in both men and women. Among non-pregnant adults, rates of serious disease range from 4.1 to 7.2 cases per 100,000 and increase with age. The average death rate for invasive infections is 8-10% for adults between ages 18-64 and 15-25% for adults>65 years of age. Serious disease is most common among elderly, bedridden patients and people suffering from severe medical conditions including diabetes mellitus, liver disease, history of stroke, history of cancer or bedsores.
Currently, disease management fully relies on antibiotics, mainly Penicillin G. In order to prevent invasive disease in newborns, pregnant women are screened for carriage of GBS at 35th to 37th weeks of gestation. Colonized mothers are then treated with high dose antibiotics during delivery to prevent neonatal GBS disease.
Current standard treatment of GBS infections is also based on antibiotics. Route, dosage, schedule and duration of therapy depend on the severity of the illness. Ten days of treatment is recommended for bacteraemia, pneumonia and soft tissue infections, while 2-3 weeks is recommended for meningitis and 3-4 weeks for osteomyelitis.
Invasive GBS diseases are associated with 5% mortality and 20% permanent damage in spite of effective antibiotic therapy, due to a very rapid and dramatic clinical course. Before prevention direct medical costs of neonatal disease were  ̃$300 million annually in the US; and GBS still poses a considerable economic burden.
Although intrapartum prophylaxis has decreased the incidence of early-onset GBS disease, currently available strategies are not ideal as they can neither prevent late-onset infections nor disease in premature babies which are at highest risk for invasive disease.
Currently, no effective preventive vaccine is available. There are efforts focusing on using capsular polysaccharides (with or without protein-conjugation) as immunogens, but several arguments militate against that approach. Polysaccharides induce IgG2 antibodies, which cross the placenta less efficiently than IgG1 or IgG3 antibodies. This especially poses a problem for the most susceptible early-born neonates, since placental antibody transfer is low before the 34th pregnancy week and about 10% of deliveries occur before that time. An additional disadvantage of polysaccharide vaccines is the incomplete vaccine coverage among GBS serotypes. Given adequate ecological pressure, replacement disease by non-vaccine serotypes remains a real threat, particularly in areas with high disease burden.
Taking these insufficiencies into account, new generation immune interventions against GBS disease are needed. Given the very recent acceptance of the use of a cervical cancer-preventing vaccine in teenage girls, a new approach would be the use of combinations of proteins as a prophylactic GBS vaccine in order to provide protection against more than one S. agalactiae strain or serotype.
Accordingly, one problem underlying the present invention was to provide alternative means for the development of medicaments such as vaccines against S. agalactiae infection. More particularly, one problem was to provide combinations of protective proteins, particularly more effective combinations, derived from S. agalactiae that can be used for the manufacture of said medicaments.
Surprisingly, this object has been solved by combinations of protective proteins/peptides comprising or consisting of the amino acid sequences as defined in SEQ ID NOS: 1 to 6 or functionally active variants thereof.
Accordingly, a first subject of the present invention relates to a composition comprising at least two proteins selected from the group consisting of

    i) a protective protein comprising or consisting of the protective peptide of SEQ ID NO: 1 (gbs0233p) or functionally active variant thereof;
    ii) a protective protein comprising or consisting of the protective peptide of SEQ ID NO: 2 (gbs1087p) or functionally active variant thereof;
    iii) a protective protein comprising or consisting of the protective peptide of SEQ ID NO: 3 (gbs1309p) or functionally active variant thereof;
    iv) a protective protein comprising or consisting of the protective peptide of SEQ ID NO: 4 (gbs1477p) or functionally active variant thereof;
    v) a protective protein comprising or consisting of the protective peptide of SEQ ID NO: 5 (gbs1478p) or functionally active variant thereof; and
    vi) a protective protein comprising or consisting of the protective peptide of SEQ ID NO: 6 (gbs2018p) or functionally active variant thereof.


Surprisingly, it was found that combinations of the above protective proteins provide a better protection against S. agalactiae than a protective protein when used alone. A better protection in the context of the present invention may refer to a situation in which protection provided by the combination is improved quantitatively in comparison to the single components of the composition. For example, the combination may provide protection against at least one serotype of S. agalactiae against which at least one of the protective proteins present in the composition does not provide protection. Accordingly, the number of serotypes against which the combination provides protection is increased. Additionally or alternatively, protection provided by the combination is improved qualitatively in comparison to the single components of the composition. For example, the survival of mice challenged with GBS strains may be improved when a composition of protective proteins is used in comparison to the single components of the composition. Both, quantitatively and qualitatively sufficient protection, are important for successful prevention and/or treatment, since it is the goal striven for to provide protection which is as high as possible and which protects against as many serotypes as possible.
Additionally, combinations of different protective proteins are in general advantageous in comparison to single protective proteins, since in the case of vaccines employing different protective proteins/antibodies the probability of a serotype switch of the pathogen in question leading to reduced effectiveness of the vaccine is strongly diminished. This is due to the fact that more than one mutation in S. agalactiae proteins at defined sites would be required in order to render the respective S. agalactiae strain unsusceptible to the vaccine.
The protective protein consisting of the amino acid sequence of SEQ ID NO: 1 is derived from S. agalactiae strain 12403 and has been denoted by gbs0233p (partial gbs0233) in accordance with the genome of NEM316 (ATCC12403). The DNA sequence encoding the full length protein gbs0233 (consisting of 308 amino acids; SEQ ID NO: 229) from which the protective protein consisting of the amino acid sequence of the SEQ ID NO: 1 is derived is disclosed at GenBank® accession number AL732656 (complete genome of Streptococcus agalactiae NEM316) and the amino acid sequence of the full length protein is disclosed in WO2004/099242 (see SEQ ID NO: 475). The amino acid sequence of SEQ ID NO: 1 is disclosed in the Examples as well as in the attached Sequence listing. The protective protein comprising or consisting of the protective peptide of SEQ ID NO: 1 or a functionally active variant thereof are referred to as (protective) proteins of subgroup i).
The protective protein consisting of the amino acid sequence of SEQ ID NO: 2 is derived from S. agalactiae strain 6313 and has been denoted by gbs1087p (partial gbs1087) in accordance with the genome of NEM316 (ATCC12403). The amino acid and encoding DNA sequences of the full length protein gbs1087 (also referred to as FbsA and consisting of 442 amino acids; SEQ ID NO: 230) from which the protective protein consisting of the amino acid sequence of the SEQ ID NO: 2 is derived is disclosed in WO2004/035618 (see FIG. 1 and SEQ ID NO: 11). The amino acid sequence of SEQ ID NO: 2 is disclosed in the Examples as well as in the attached Sequence listing. The protective protein comprising or consisting of the protective peptide of SEQ ID NO: 2 or a functionally active variant thereof are referred to as (protective) proteins of subgroup ii).
The protective protein consisting of the amino acid sequence of SEQ ID NO: 3 is derived from S. agalactiae strain 12403 and has been denoted by gbs1309p (partial gbs1309) in accordance with the genome of NEM316 (ATCC12403). The DNA sequence encoding the full length protein gbs1309 (consisting of 403 amino acids; SEQ ID NO: 231) from which the protective protein consisting of the amino acid sequence of the SEQ ID NO: 3 is derived is disclosed in GenBank® accession number AL732656 (complete genome of Streptococcus agalactiae NEM316) and the amino acid sequence of the full length protein is disclosed in WO2004/099242 (see SEQ ID NO: 307). The amino acid sequence of SEQ ID NO: 3 is disclosed in the Examples as well as in the attached Sequence listing. The protective protein comprising or consisting of the protective peptide of SEQ ID NO: 3 or a functionally active variant thereof are referred to as (protective) proteins of subgroup iii).
The protective protein consisting of the amino acid sequence of SEQ ID NO: 4 is derived from S. agalactiae strain 6313 and has been denoted by gbs1477p (partial gbs1477) in accordance with the genome of NEM316 (ATCC12403). The amino acid and encoding DNA sequences of the full length protein gbs1477 (also referred to as PabB and consisting of 674 amino acids; SEQ ID NO: 232) from which the protective protein consisting of the amino acid sequence of the SEQ ID NO: 4 is derived is disclosed in WO2004/035618 (see FIG. 16 and SEQ ID NO: 18). The amino acid sequence of SEQ ID NO: 4 is disclosed in the Examples as well as in the attached Sequence listing. The protective protein comprising or consisting of the protective peptide of SEQ ID NO: 4 or a functionally active variant thereof are referred to as (protective) proteins of subgroup iv).
The protective protein consisting of the amino acid sequence of SEQ ID NO: 5 is derived from S. agalactiae strain 6313 and has been denoted by gbs1478p (partial gbs1478) in accordance with the genome of NEM316 (ATCC12403). The amino acid and encoding DNA sequences of the full length protein gbs1478 (also referred to as PabA and consisting of 901 amino acids; SEQ ID NO: 233) from which the protective protein consisting of the amino acid sequence of the SEQ ID NO: 5 is derived is disclosed in WO2004/035618 (see FIG. 16 and SEQ ID NO: 17). The amino acid sequence of SEQ ID NO: 5 is disclosed in the Examples as well as in the attached Sequence listing. The protective protein comprising or consisting of the protective peptide of SEQ ID NO: 5 or a functionally active variant thereof are referred to as (protective) proteins of subgroup v).
The protective protein consisting of the amino acid sequence of SEQ ID NO: 6 is derived from S. agalactiae strain 12403 and has been denoted by gbs2018p (partial gbs2018) in accordance with the genome of NEM316 (ATCC12403). The DNA sequence encoding the full length protein gbs2018 (also referred to as BibA (Santi et al., 2007, Mol. Microbiol. 63:754-767) and consisting of 643 amino acids; SEQ ID NO: 234) from which the protective protein consisting of the amino acid sequence of the SEQ ID NO: 6 is derived is disclosed at GenBank® accession number AL732656 (complete genome of Streptococcus agalactiae NEM316) and the amino acid sequence of the full length protein is disclosed in WO2004/099242 (see SEQ ID NO: 364). The amino acid sequence of SEQ ID NO: 6 is disclosed in the Examples as well as in the attached Sequence listing. The protective protein comprising or consisting of the protective peptide of SEQ ID NO: 6 or a functionally active variant thereof are referred to as (protective) proteins of subgroup vi).
The combinations of the protective proteins of the sequences of SEQ ID NO: 1 to 6 have been shown to induce a protective immune response against different serotypes and/or to show increased protection against S. agalactiae in an animal model (see Examples). Functionally active variants may be obtained by changing the sequence of at least one of the protective proteins of SEQ ID NO: 1 to 6 and are characterized by having a biological activity similar to that displayed by the respective protective protein of the sequence of SEQ ID NO: 1 to 6 from which the variant is derived, including the ability to induce protective immune responses and/or to show protection against S. agalactiae e.g. in an animal model, wherein any variant may be tested in any of the tests described in the Examples. The functionally active variant of a protective protein may be obtained by sequence alterations in the protective protein, wherein the protein with the sequence alterations essentially retains a function of the unaltered protective protein, e.g. having a biological activity similar to that displayed by the unaltered protective protein (see above) including the ability to induce protective immune responses and/or to show protection against S. agalactiae. Such sequence alterations can include, but are not limited to, (conservative) substitutions, deletions, mutations and insertions.
In a preferred embodiment of the invention the composition comprises at least three proteins selected from the group consisting of subgroup i) to vi). In an even more preferred embodiment of the invention the composition comprises at least four proteins selected from the group consisting of subgroup i) to vi).
In a preferred embodiment of the invention the at least two, three or four proteins of the composition of the invention are selected from different subgroups i) to vi). Alternatively or additionally, at least two of the proteins of the composition of the invention are selected from one of the subgroups i) to vi).
Examples of combinations of the first alternative (selection of protective proteins from different groups) are compositions comprising:

    
    
        one protein of subgroup i) and one protein of subgroup ii);
        one protein of subgroup i) and one protein of subgroup iii);
        one protein of subgroup i) and one protein of subgroup iv);
        one protein of subgroup i) and one protein of subgroup v);
        one protein of subgroup i) and one protein of subgroup vi);
        one protein of subgroup ii) and one protein of subgroup iii);
        one protein of subgroup ii) and one protein of subgroup iv);
        one protein of subgroup ii) and one protein of subgroup v);
        one protein of subgroup ii) and one protein of subgroup vi);
        one protein of subgroup iii) and one protein of subgroup iv);
        one protein of subgroup iii) and one protein of subgroup v);
        one protein of subgroup iii) and one protein of subgroup vi);
        one protein of subgroup iv) and one protein of subgroup v);
        one protein of subgroup iv) and one protein of subgroup vi);
        one protein of subgroup v) and one protein of subgroup vi);
        one protein of subgroup i) and one protein of subgroup ii) and one protein selected from any of the subgroups iii) to vi);
        one protein of subgroup i) and one protein of subgroup iii) and one protein selected from any of the subgroups ii) or iv) to vi);
        one protein of subgroup i) and one protein of subgroup iv) and one protein selected from any of the subgroups ii), iii), v) or vi);
        one protein of subgroup i) and one protein of subgroup v) and one protein selected from any of the subgroups ii) to iv) or vi);
        one protein of subgroup i) and one protein of subgroup vi) and one protein selected from any of the subgroups ii) to v);
        one protein of subgroup ii) and one protein of subgroup iii) and one protein selected from any of the subgroups i) or iv) to vi);
        one protein of subgroup ii) and one protein of subgroup iv) and one protein selected to from any of the subgroups i), iii), v) or vi);
        one protein of subgroup ii) and one protein of subgroup v) and one protein selected from any of the subgroups i), iii), iv) or vi);
        one protein of subgroup ii) and one protein of subgroup vi) and one protein selected from any of the subgroups i) or iii) to v);
        one protein of subgroup iii) and one protein of subgroup iv) and one protein selected from any of the subgroups i), ii), v) or vi);
        one protein of subgroup iii) and one protein of subgroup v) and one protein selected from any of the subgroups i), ii), iv) or vi);
        one protein of subgroup iii) and one protein of subgroup vi) and one protein selected from any of the subgroups i), ii), iv) or v);
        one protein of subgroup iv) and one protein of subgroup v) and one protein selected from any of the subgroups i) to iii) or vi);
        one protein of subgroup iv) and one protein of subgroup vi) and one protein selected from any of the subgroups i) to iii) or v); or
        one protein of subgroup v) and one protein of subgroup vi) and one protein selected from any of the subgroups i) to iv).
    
    


Preferred examples are:

    
    
        one protein of subgroup iv) and one protein of subgroup vi);
        one protein of subgroup iv), one protein of subgroup vi) and one protein of subgroup ii);
        one protein of subgroup iv), one protein of subgroup vi), one protein of subgroup ii) and one protein of subgroup v);
        one protein of subgroup iv), one protein of subgroup vi), one protein of subgroup ii) one protein of subgroup v) and one protein of subgroup i);
        one protein of subgroup iv), one protein of subgroup vi), one protein of subgroup ii), one protein of subgroup v) and one protein of subgroup iii); or
        one protein of subgroup iv), one protein of subgroup vi), one protein of subgroup ii), one protein of subgroup v), one protein of subgroup i) and one protein of subgroup iii).
    
    


In an alternative preferred embodiment of the invention at least two of the proteins of the composition of the invention may be selected from one of the subgroups i) to vi). The at least two proteins may be selected in order to cover different strains or serotypes of S. agalactiae and, accordingly, to provide protection against e.g. different strains or serotypes of S. agalactiae. The complete genome of Streptococcus agalactiae NEM316 (strain 12403) is available at GenBank® accession number AL732656. Furthermore, the complete or incomplete genomic sequences of the following strains of Streptococcus agalactiae are available at GenBank® (NIH genetic sequence database; http://www.ncbi.nlm.nih.gov/) or NCBI (National Center for Biotechnology Information, Bethesda, Md., USA; http://www.ncbi.nlm.nih.gov/) using the indicated accession numbers:















Strain
Serotype
Source









515
Ia
NCBI: NZ_AAJP00000000





GenBank ®: AAJP00000000



A909
Ia/c
NCBI: NC_007432





GenBank ®: CP000114



H36B
Ib
NCBI: NZ_AAJS00000000





GenBank ®: AAJS00000000



18RS21
II
NCBI NZ_AAJO00000000





GenBank ®: AAJO00000000



COH1
III
NCBI: NZ_AAJR00000000





GenBank ®: AAJR00000000



ATCC12403
III
NCBI: NC_004368



(NEM316)

GenBank ®: AL732656



2603V/R
V
NCBI: NC_004116





GenBank ®: AE009948



CJB111
V
NCBI: NZ_AAJQ00000000





GenBank ®: AAJQ00000000










Using the sequences of SEQ ID NO: 1 to 6 as specified in the present description and knowing the sequences of other S. agalactiae strains (e.g. vide supra) the skilled person is able to identify the corresponding sequences of S. agalactiae strains other than 12403 (for SEQ ID NO: 1, 3 and 6) or 6313 (for SEQ ID NO: 2, 4 and 5) without undue burden. The corresponding sequences may be identified using e.g. the tools and sequences provided by “The Comprehensive Microbial Resource (CMR)” (see http://cmr.tigr.org/). However, it should be understood that the above strains are listed as examples of different S. agalactiae strains and that the present invention is not to be limited to those strains.
Additionally, examples of sequences of the proteins corresponding to SEQ ID NO: 1 to 6 and derived from other serotypes are published or disclosed in:











Protein or



Analogue
NCBI Accession Number (strain [serotype]) or SEQ ID NO







gbs0233
Any sequence of SEQ ID NO: 55 to 60 (see Table 7),



229 and 235 to 286.


gbs1087
CAD12883 (6313); CAD27183 (706 S2 [Ia]); CAD27181



(SS1169 [V]); CAD27186 (O90R); CAD27182 (O176



H4A [II]) or any sequence of SEQ ID NO: 67 to 72



(see Table 8), 230 and 287 to 316.


gbs1309
Any sequence of SEQ ID NO: 79 to 84 (see Table 9),



231 and 317 to 359.


gbs1477
Any sequence of SEQ ID NO: 91 to 132 (see Table 10),



223 to 228 (see FIG. 5), 232 and 360 to 362.


gbs1478
Any sequence of SEQ ID NO: 185 to 203 (see Table 11),



233 and 363 to 378.


gbs2018
CAJ66802 (CCH57); CAJ66794 (CCH180); CAJ66788



(NEM1002); CAJ 66790 (NEM1560) or any sequence of SEQ



ID NO: 175 to 179 (see Table 12), 234 and 379 to 425.









However, it should be understood that the present invention is not limited to the variants and corresponding proteins described above. Other naturally occurring proteins corresponding to those of SEQ ID NO: 1 to 6 may be identified as described above and used in order to carry out the present invention.
Examples of combinations of the second alternative (selection of protective proteins from one group only) are compositions comprising:

    
    
        at least two different proteins of subgroup i), preferably selected from the group consisting of proteins comprising or consisting of SEQ ID NO: 1 or a naturally occurring variant thereof;
        at least two different proteins of subgroup ii), preferably selected from the group consisting of proteins comprising or consisting of SEQ ID NO: 2 or a naturally occurring variant thereof;
        at least two different proteins of subgroup iii), preferably selected from the group consisting of proteins comprising or consisting of SEQ ID NO: 3 or a naturally occurring variant thereof;
        at least two different proteins of subgroup iv), preferably selected from the group consisting of proteins comprising or consisting of SEQ ID NO: 4 or a naturally occurring variant thereof;
        at least two different proteins of subgroup v), preferably selected from the group consisting of proteins comprising or consisting of SEQ ID NO: 5 or a naturally occurring variant thereof;
        at least two different proteins of subgroup vi), preferably selected from the group consisting of proteins comprising or consisting of SEQ ID NO: 6 or a naturally occurring variant thereof;
        preferably at least two protective proteins each comprising or consisting of a sequence selected from SEQ ID NO: 55 to 60 (see Table 7), 235 to 286, and optionally 229;
        preferably at least two protective proteins each comprising or consisting of a sequence selected from SEQ ID NO: 67 to 72 (see Table 8), 287 to 316, and optionally 230;
        preferably at least two protective proteins each comprising or consisting of a sequence selected from SEQ ID NO: 79 to 84 (see Table 9), 317 to 359, and optionally 231;
        preferably at least two protective proteins each comprising or consisting of a sequence selected from SEQ ID NO: 185 to 203 (see Table 11), 363 to 378, and optionally 233;
        preferably at least two protective proteins each comprising or consisting of a sequence selected from SEQ ID NO: 175 to 179 (see Table 12), 379 to 425 and optionally 234; or
        more preferably at least two protective proteins each comprising or consisting of a sequence selected from SEQ ID NO: 91 to 132 (see Table 10), 360 to 362, or 223 to 228 (see FIG. 5) and optionally 232.
    
    


In a preferred embodiment the naturally occurring variants are those derived from S. agalactiae strains selected from the group consisting of IC97, IC98, IC105, IC108, IC216, IC244, IC245, IC246, IC247, IC250, IC251, IC252, IC253, IC254, IC255, IC287, IC288, IC289, IC290, IC291, IC304, IC305, IC306, IC361, IC363, IC364, IC365, IC366, IC367, IC368, IC377, IC379, IC432, IC434, IC455, IC457, IC458, IC459, IC460, IC461, IC462, IC463, IC469, IC470, 126H4A, 5095S2, 6313, 12351, 12403 (NEM316), 12401, COH1, BAA23, 0176H4A, A909, C388/90, BAA22, 2603V/R, 49447, BAA611, 515, H36B, 18RS21, CJB111, and those disclosed in Tables 7 to 13.
In another preferred embodiment of the present invention the composition of the invention comprises

    
    
        at least one protein of subgroup iv);
        at least one protein of subgroup vi);
        at least one protein of subgroup iv) and at least one protein of subgroup vi);
        at least one protein of subgroup iv), at least one protein of subgroup vi) and at least one protein of subgroup ii); or
        at least one protein of subgroup iv), at least one protein of subgroup vi), at least one protein of subgroup ii) and at least one protein of subgroup v).
    
    


In a preferred embodiment of the invention one of the at least two proteins comprises or consists of the protective peptide of SEQ ID NO: 4 (gbs1477p) or a functionally active variant thereof, preferably protective peptide of SEQ ID NO: 4 (gbs 1477p) or a naturally occurring functionally active variant thereof, more preferably a naturally occurring functionally active variant as listed in Tables 10, 13, and in the Sequence listing.
In another preferred embodiment of the invention one of the at least two proteins comprises or consists of the protective peptide of SEQ ID NO: 6 (gbs2018p) or a functionally active variant thereof, preferably protective peptide of SEQ ID NO: 6 (gbs2018p) or a naturally occurring functionally active variant thereof, more preferably a naturally occurring functionally active variant as listed in Tables 12, 13, and in the Sequence listing.
In another preferred embodiment of the invention one of the at least two proteins comprises or consists of the protective peptide of SEQ ID NO: 2 (gbs1087p) or a functionally active variant thereof, preferably protective peptide of SEQ ID NO: 2 (gbs1087p) or a naturally occurring functionally active variant thereof, more preferably a naturally occurring functionally active variant as listed in Tables 8, 13, and in the Sequence listing.
In another preferred embodiment of the invention one of the at least two proteins comprises or consists of the protective peptide of SEQ ID NO: 5 (gbs1478p) or a functionally active variant thereof, preferably protective peptide of SEQ ID NO: 5 (gbs1478p) or a naturally occurring functionally active variant thereof, more preferably a naturally occurring functionally active variant as listed in Tables 11, 13, and in the Sequence listing.
In another preferred embodiment of the invention one of the at least two proteins comprises or consists of the protective peptide of SEQ ID NO: 1 (gbs0233p) or a functionally active variant thereof, preferably protective peptide of SEQ ID NO: 1 (gbs0233p) or a naturally occurring functionally active variant thereof, more preferably a naturally occurring functionally active variant as listed in Tables 7, 13, and in the Sequence listing.
In another preferred embodiment of the invention one of the at least two proteins comprises or consists of the protective peptide of SEQ ID NO: 3 (gbs1309p) or a functionally active variant thereof, preferably protective peptide of SEQ ID NO: 3 (gbs1309p) or a naturally occurring functionally active variant thereof, more preferably a naturally occurring functionally active variant as listed in Tables 9, 13, and in the Sequence listing.
In a more referred embodiment of the invention the at least two proteins of the composition of the invention encompass:

    
    
        the protective peptide of SEQ ID NO: 4 (gbs1477p) and the protective peptide of SEQ ID NO: 6 (gbs2018p);
        the protective peptide of SEQ ID NO: 4 (gbs1477p) and the protective peptide of SEQ ID NO: 6 (gbs2018p) and the protective peptide of SEQ ID NO: 2 (gbs1087p); or
        the protective peptide of SEQ ID NO: 4 (gbs1477p) and the protective peptide of SEQ ID NO: 6 (gbs2018p) and the protective peptide of SEQ ID NO: 2 (gbs1087p) and the protective peptide of SEQ ID NO: 5 (gbs1478p).
    
    


In a further preferred embodiment of the invention a naturally occurring functionally active variant of any of the protective peptides of SEQ ID NO: 1 to 6 of the above list of compositions may be used. Examples of the resulting combinations are:

    I. the protective peptide of SEQ ID NO: 4 (gbs1477p) and a naturally occurring functionally active variant of SEQ ID NO: 6 (gbs2018p);
    II. a naturally occurring functionally active variant of SEQ ID NO: 4 (gbs1477p) and the protective peptide of SEQ ID NO: 6 (gbs2018p);
    III. a naturally occurring functionally active variant of SEQ ID NO: 4 (gbs1477p) and a naturally occurring functionally active variant of SEQ ID NO: 6 (gbs2018p);
    IV. the protective peptide of SEQ ID NO: 4 (gbs1477p) and the protective peptide of SEQ ID NO: 6 (gbs2018p) and a naturally occurring functionally active variant of SEQ ID NO: 2 (gbs1087p);
    V. the protective peptide of SEQ ID NO: 4 (gbs1477p) and a naturally occurring functionally active variant of SEQ ID NO: 6 (gbs2018p) and the protective peptide of SEQ ID NO: 2 (gbs1087p);
    VI. a naturally occurring functionally active variant of SEQ ID NO: 4 (gbs1477p) and the protective peptide of SEQ ID NO: 6 (gbs2018p) and the protective peptide of SEQ ID NO: 2 (gbs1087p);
    VII. a naturally occurring functionally active variant of SEQ ID NO: 4 (gbs1477p) and a naturally occurring functionally active variant of SEQ ID NO: 6 (gbs2018p) and the protective peptide of SEQ ID NO: 2 (gbs1087p);
    VIII. the protective peptide of SEQ ID NO: 4 (gbs1477p) and a naturally occurring functionally active variant of SEQ ID NO: 6 (gbs2018p) and a naturally occurring functionally active variant of SEQ ID NO: 2 (gbs1087p);
    IX. a naturally occurring functionally active variant of SEQ ID NO: 4 (gbs1477p) and the protective peptide of SEQ ID NO: 6 (gbs2018p) and a naturally occurring functionally active variant of SEQ ID NO: 2 (gbs1087p);
    X. a naturally occurring functionally active variant of SEQ ID NO: 4 (gbs1477p) and a naturally occurring functionally active variant of SEQ ID NO: 6 (gbs2018p) and a naturally occurring functionally active variant of SEQ ID NO: 2 (gbs1087p);
    XI. any of the compositions of I to X in combination with the protective peptide of SEQ ID NO: 5 (gbs1478p); or
    XII. any of the compositions of I to X in combination with a naturally occurring functionally active variant of SEQ ID NO: 5 (gbs1478p).

wherein the naturally occurring functionally active variant is selected form those listed in Tables 7 to 13 and FIG. 5 and those of SEQ ID NO: 229 to 234 and 235 to 425.



Preferred S. agalactiae strains from which the naturally occurring functionally active variant may be derived include IC97, IC98, IC105, IC108, IC216, IC244, IC245, IC246, IC247, IC250, IC251, IC252, IC253, IC254, IC255, IC287, IC288, IC289, IC290, IC291, IC304, IC305, IC306, IC361, IC363, IC364, IC365, IC366, IC367, IC368, IC377, IC379, IC432, IC434, IC455, IC457, IC458, IC459, IC460, IC461, IC462, IC463, IC469, IC470, 126H4A, 5095S2, 6313, 12351, 12403 (NEM316), 12401, COH1, BAA23, 0176H4A, A909, C388/90, BAA22, 2603V/R, 49447, BAA611, 515, H36B, 18RS21, CJB111, and those disclosed in Tables 7 to 13.
In one embodiment of the invention two or more proteins of the at least two proteins of the composition of the invention may be combined into at least one fusion protein. The resulting fusion protein may encompass two or more of the proteins of subgroups i), ii), iii), iv), v) and/or vi) as defined above. Any of the specific combinations mentioned above may be combined into at least one fusion protein. The fusion protein may encompass e.g. the same protein components as described in the section “examples of combinations of the first alternative (selection of protective proteins from different groups)” for the components of the composition, wherein the proteins can be arranged in the fusion protein in any suitable manner.
The fusion protein may comprise or consist of two or more proteins as defined above. Additionally, the fusion protein may encompass a linker, such as a protein linker, to connect the two or more proteins or additional C- or N-terminal sequences, such as a tag in order to purify the fusion protein. Additional sequences may also result from genetic engineering and the use of suitable restriction sites when preparing the nucleic acid sequences underlying the fusion protein.
The proteins of subgroup i), ii), iii), iv), v) and/or vi) combined in a fusion protein may be directly joined to each other or may be combined over a linker. The linker may be e.g. a short amino acid sequence. The linker may result from the genetic engineering of a suitable fusion protein or may be introduced in order to allow the single proteins to operate effectively.
In a preferred embodiment the functionally active variant

    a) is a functionally active fragment of the protective peptide, the functionally active fragment comprising at least 50% of the sequence of the protective peptide, preferably at least 70%, more preferably at least 80%, still more preferably at least 90%, even more preferably at least 95% and most preferably at least 97%, 98% or 99%;
    b) is derived from the protective peptide by at least one amino acid substitution, addition and/or deletion, wherein the functionally active variant has a sequence identity to the protective peptide or to the functionally active fragment as defined in a) of at least 40%, preferably at least 60%, more preferably at least 75%, still more preferably at least 90%, even more preferably at least 95% and most preferably at least 97%, 98% or 99%; and/or
    c) consists of the protective peptide or a functionally active variant thereof and additionally at least one amino acid heterologous to the protective peptide,

preferably wherein the functionally active variant is derived from or identical to any of the naturally occurring variants of any of the sequences of SEQ ID NO: 55 to 60, 67 to 72, 79 to 84, 91 to 132, 175 to 179, 185 to 203, 223 to 234, and 235 to 425.



The combinations of protective peptides of SEQ ID NO: 1 to 6 have been shown to induce a protective immune response against different serotypes and/or to show protection against S. agalactiae in a sepsis model (see Examples). Functionally active variants may be obtained by changing the sequence of the protective peptide as defined above and are characterized by having a biological activity similar to that displayed by the respective protective peptide of the sequence of SEQ ID NO: 1 to 6 from which the variant is derived, including the ability to induce protective immune responses and/or to show protection against S. agalactiae e.g. in a sepsis model, wherein any variant may be tested in any of the tests described in the Examples within a composition of at least two proteins as defined above. The functionally active variant of a protective peptide may be obtained by sequence alterations in the protective peptide, wherein the peptide with the sequence alterations retains a function of the unaltered protective peptide, e.g. having a biological activity similar to that displayed by the unaltered protective peptide (see above), when used in combination of the invention. Such sequence alterations can include, but are not limited to, (conservative) substitutions, additions, deletions, mutations and insertions.
The variant of the protective peptide is functionally active in the context of the present invention, if the activity of the composition of the invention including the variant (but not the original protein) amounts to at least 10%, preferably at least 25%, more preferably at least 50%, even more preferably at least 70%, still more preferably at least 80%, especially at least 90%, particularly at least 95%, most preferably at least 99% of the activity of the composition of the invention including the protective peptide without sequence alteration (i.e. the original protein). The activity of the composition including the variant may be determined or measured as described in the Examples and then compared to that obtained for the composition including the respective protective peptide of the amino acid sequence of SEQ ID NO: 1 to 6 instead of the variant.
The functionally active fragment of the protective peptide is characterized by being derived from the protective peptide of SEQ ID NO: 1 to 6 by one or more deletions resulting in a peptide comprising at least 50% of the sequence of the protective peptide, preferably at least 70%, more preferably at least 80%, still more preferably at least 90%, even more preferably at least 95% and most preferably at least 97%, 98% or 99%. Sequence identity may be determined as described below. The deletion(s) may be C-terminally, N-terminally and/or internally. Preferably the fragment is obtained by 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, more preferably 1, 2, 3, 4 or 5, even more preferably 1, 2 or 3, still more preferably 1 or 2, most preferably 1 deletion(s).
Alternatively or additionally, the variant may be obtained from the protective peptide by at least one amino acid substitution, addition and/or deletion, wherein the functionally active variant has a sequence identity to the protective peptide or to the functionally active fragment as defined in a) of at least 40%, preferably at least 60%, more preferably at least 75%, still more preferably at least 90%, even more preferably at least 95% and most preferably at least 97%, 98% or 99%. Sequence identity may be determined as described below. The substitution(s), addition(s) and/or deletion(s) may be C-terminally, N-terminally and/or internally. Preferably, the functionally active variant is obtained from the protective peptide or the fragment, preferably the protective peptide, by 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, more preferably 1, 2, 3, 4 or 5, even more preferably 1, 2 or 3, still more preferably 1 or 2, most preferably 1 amino acid substitution(s), addition(s) and/or deletion(s).
Furthermore, the variant may consist of the protective peptide or the functionally active variant thereof, preferably the variant of a) and/or b), and at least one amino acid residue heterologous to the protective peptide or variant thereof, such as a marker protein. The feature “heterologous amino acid” or “amino acid heterologous to the protective peptide or variant thereof” refers to any amino acid which is different from that amino acid located adjacent to the protective protein in any naturally occurring protein of S. agalactiae, particularly from that of strain 12403 (for SEQ ID NO: 1, 3 and 6) or 6313 (for SEQ ID NO: 2, 4 and 5), especially the sequence made reference to above. The one or more additional amino acids may be C-terminally, N-terminally or C- and N-terminally to the protective peptide or variant thereof.
The substituted or additional sequence or amino acid residue(s) as defined above consists of (an) amino acid residue(s), which may be any amino acid, which may be either an L- and/or a D-amino acid, naturally occurring and otherwise. Preferably the amino acid is any naturally occurring amino acid such as alanine, cysteine, aspartic acid, glutamic acid, phenylalanine, glycine, histidine, isoleucine, lysine, leucine, methionine, asparagine, proline, glutamine, arginine, serine, threonine, valine, tryptophan or tyrosine.
However, the amino acid may also be a modified or an unusual amino acid. Examples of those are 2-aminoadipic acid, 3-aminoadipic acid, beta-alanine, 2-aminobutyric acid, 4-aminobutyric acid, 6-aminocaproic acid, 2-aminoheptanoic acid, 2-aminoisobutyric acid, 3-aminoisobutyric acid, 2-aminopimelic acid, 2,4-diaminobutyric acid, desmosine, 2,2′-diaminopimelic acid, 2,3-diaminopropionic acid, N-ethylglycine, N-ethylasparagine, hydroxylysine, allo-hydroxylysine, 3-hydroxyproline, 4-hydroxyproline, isodesmosine, allo-isoleucine, N-methylglycine, N-methylisoleucine, 6-N-methyllysine, N-methylvaline, norvaline, norleucine or ornithine. Additionally, the amino acid may be subject to modifications such as posttranslational modifications. Examples of modifications include acetylation, amidation, blocking, formylation, gamma-carboxyglutamic acid hydroxylation, glycosylation, methylation, phosphorylation and sulfatation. If more than one substituted or additional heterologous amino acid residue is present in the peptide, the amino acid residues may be the same or different from one another.
In one preferred embodiment of the invention, the functionally active variant of the peptide of the invention is essentially identical to the protective peptide of subgroups i) to vi), but differs from the peptide of the SEQ ID NO: 1 to 6, respectively, in that it is derived from a homologous sequence of a different strain or even serotype of S. agalactiae. As detailed above different strains and serotypes of S. agalactiae have been identified so far. Accordingly, any of these serotypes may be the basis for the functionally active variant. These are referred to as naturally occurring variants (see also above). Preferably, these naturally occurring variants are derived from S. agalactiae strains selected from the group consisting of IC97, IC98, IC105, IC108, IC216, IC244, IC245, IC246, IC247, IC250, IC251, IC252, IC253, IC254, IC255, IC287, IC288, IC289, IC290, IC291, IC304, IC305, IC306, IC361, IC363, IC364, IC365, IC366, IC367, IC368, IC377, IC379, IC432, IC434, IC455, IC457, IC458, IC459, IC460, IC461, IC462, IC463, IC469, IC470, 126H4A, 5095S2, 6313, 12351, 12403 (NEM316), 12401, COH1, BAA23, 0176H4A, A909, C388/90, BAA22, 2603V/R, 49447, BAA611, 515, H36B, 18RS21, CJB111, and those disclosed in Tables 7 to 13.
However, the term “functionally active variant” includes naturally occurring allelic variants, as well as mutants or any other non-naturally occurring variants. As is known in the art, an allelic variant is an alternate form of a (poly)peptide that is characterized as having a substitution, deletion, or addition of one or more amino acids that does essentially not alter the biological function of the polypeptide. By “biological function” is meant a function of the peptide in the cell it naturally occurs in, even if the function is not necessary for the growth or survival of the cells. For example, the biological function of a porin is to allow the entry into cells of compounds present in the extracellular medium. The biological function is distinct from the antigenic function. A polypeptide can have more than one biological function.
Accordingly, the present invention also relates to compositions comprising protective peptides including functionally active variants thereof of different S. agalactiae isolates. Such homologues may easily be identified and isolated based on the nucleic acid and amino acid sequences disclosed herein as discussed above. A homologous protective peptide of a different strain or even serotype may be identified by e.g. sequence alignment. The homologous sequence may vary from any of the protective peptides of subgroups i) to vi), by one or more amino acid substitutions, deletions and/or additions.
Percentage of sequence identity can be determined e.g. by sequence alignment. Methods of alignment of sequences for comparison are well known in the art. Various programs and alignment algorithms have been described e.g. in Smith and Waterman, Adv. Appl. Math. 2: 482, 1981 or Pearson and Lipman, Proc. Natl. Acad. Sci. U.S.A. 85: 2444-2448, 1988.
The NCBI Basic Local Alignment Search Tool (NCBI BLAST) (Altschul et al., J. Mol. Biol. 215: 403-410, 1990) is available from several sources, including the National Center for Biotechnology Information (NCBI, Bethesda, Md.) and on the Internet, for use in connection with the sequence analysis programs blastp, blastn, blastx, tblastn and tblastx. Variants, e.g. of any protective peptide of the sequences of SEQ ID NO: 1 to 6, are typically characterized using the NCBI Blast 2.0, gapped blastp set to default parameters. For comparisons of amino acid sequences of e.g. at least 85 amino acids, the “Blast 2 sequences” function may be employed using the default BLOSUM62 matrix set to default parameters, (gap existence cost of 11, and a per residue gap cost of 1).
In a preferred embodiment, the functionally active variant derived from the peptide as defined above by amino acid exchanges, deletions or insertions may also conserve, or more preferably improve, the activity (as defined above). Furthermore, these peptides may also cover epitopes, which trigger the same or preferably an improved T cell response. These epitopes are referred to as “heteroclitic”. They have a similar or preferably greater affinity to MHC/HLA molecules, and the ability to stimulate the T cell receptors (TCR) directed to the original epitope in a similar or preferably stronger manner. Heteroclitic epitopes can be obtained by rational design i.e. taking into account the contribution of individual residues to binding to MHC/HLA as for instance described by (Rammensee, H. et al., 1999, Immunogenetics. 50: 213-219), combined with a systematic exchange of residues potentially interacting with the TCR and testing the resulting sequences with T cells directed against the original epitope. Such a design is possible for a skilled man in the art without much experimentation.
Conservative substitutions are those that take place within a family of amino acids that are related in their side chains and chemical properties. Examples of such families are amino acids with basic side chains, with acidic side chains, with non-polar aliphatic side chains, with non-polar aromatic side chains, with uncharged polar side chains, with small side chains, with large side chains etc. In one embodiment, one conservative substitution is included in the peptide. In another embodiment, two conservative substitutions or less are included in the peptide. In a further embodiment, three conservative substitutions or less are included in the peptide.
Examples of conservative amino acid substitutions include, but are not limited to, those listed below:














Original Residue
Conservative Substitutions









Ala
Ser



Arg
Lys



Asn
Gln; His



Asp
Glu



Cys
Ser



Gln
Asn



Glu
Asp



His
Asn; Gln



Ile
Leu; Val



Leu
Ile; Val



Lys
Arg; Gln; Asn



Met
Leu; Ile



Phe
Met; Leu; Tyr



Ser
Thr



Thr
Ser



Trp
Tyr



Tyr
Trp; Phe



Val
Ile; Leu










In another embodiment of the invention the peptide as defined above may be modified by a variety of chemical techniques to produce derivatives having essentially the same activity (as defined above for fragments and variants) as the modified peptides, and optionally having other desirable properties. For example, carboxylic acid groups of the protein, whether C-terminal or side chain, may be provided in the form of a salt of a pharmaceutically-acceptable cation or esterified to form an ester, or converted to an amide. Amino groups of the peptide, whether amino-terminal or side chain, may be in the form of a pharmaceutically-acceptable acid addition salt, such as the HCl, HBr, acetic, benzoic, toluene sulfonic, maleic, tartaric and other organic salts, or may be converted to an amide.
Hydroxyl groups of the peptide side chains may be converted to alkoxy or to an ester using well recognized techniques. Phenyl and phenolic rings of the peptide side chains may be substituted with one or more halogen atoms, such as fluorine, chlorine, bromine or iodine, or with alkyl, alkoxy, carboxylic acids and esters thereof, or amides of such carboxylic acids. Thiols can be protected with any one of a number of well recognized protecting groups, such as acetamide groups.
Peptides of this invention may be in combination with outer surface proteins or other proteins or antigens of other proteins. In such combination, the peptide may be in the form of a fusion protein. The peptides/proteins of the composition of the invention may be optionally fused to a selected peptide or protein derived from other microorganisms. For example, a peptide or protein may be fused at its N-terminus or C-terminus to a polypeptide from another pathogen or to more than one polypeptide in sequence. Peptides which may be useful for this purpose include polypeptides identified by the prior art.
In a preferred embodiment of the invention a protein/peptide of the composition of the invention is fused to an epitope tag which provides an epitope to which an anti-tag substance can selectively bind. The epitope tag is generally placed at the N- or C-terminus of the peptide but may be incorporated as an internal insertion or substitution as the biological activity permits. The presence of such epitope-tagged forms of a peptide can be detected using a substance such as an antibody against the tagged peptide. Also, provision of the epitope tag enables the peptide to be readily purified by affinity purification using an anti-tag antibody or another type of affinity matrix that binds to the epitope tag. Various tag polypeptides and their respective antibodies are well known in the art. Examples include a poly-histidine (poly-his) tag, e.g. a hexa-histidine tag as described in the Examples, a poly-histidine-glycine (poly-his-gly) tag, the HA tag polypeptide, the c-myc tag, the Strep tag and the FLAG tag.
Fusions also may include the peptides/proteins of the composition of this invention fused or coupled to moieties other than amino acids, including lipids and carbohydrates. Further, peptides/proteins/compositions of this invention may be employed in combination with other vaccinal agents described by the prior art, as well as with other types of vaccinal agents derived from other microorganisms. Such peptides/proteins are useful in the prevention, treatment and diagnosis of diseases caused by a wide spectrum of Streptococcus isolates.
These fusion proteins are constructed for use in the methods and compositions of this invention. These fusion proteins or multimeric proteins may be produced recombinantly, or may be synthesized chemically.
The peptides and proteins described herein may be prepared by any of a number of conventional techniques. Desired peptides may be chemically synthesized. An alternative approach involves generating the fragments of known peptides by enzymatic digestion, e.g., by treating the protein with an enzyme known to cleave proteins at sites defined by particular amino acid residues, or by digesting the DNA with suitable restriction enzymes, expressing the digested DNA and isolating the desired fragment. Yet another suitable technique involves isolating and amplifying a DNA fragment encoding a desired peptide fragment, by polymerase chain reaction (PCR). Oligonucleotides that define the desired termini of the DNA fragment are employed as the 5′ and 3′ primers in the PCR. Techniques for making mutations, such as deletions, insertions and substitutions, at predetermined sites in DNA, and therefore in proteins having a known sequence are well known. One of skill in the art using conventional techniques, such as PCR, may readily use the peptides, proteins and compositions provided herein to identify and isolate other similar proteins. Such methods are routine and not considered to require undue experimentation, given the information provided herein. For example, variations can be made using oligonucleotide-mediated site-directed mutagenesis (Carter et al., Nucl. Acids Res., 13:4431 (1985); Zoller et al., Nucl. Acids Res. 10:6487 (1987)), cassette mutagenesis (Wells et al., Gene, 34:315 (1985)), restriction selection mutagenesis (Wells et al., Philos. Trans. R. Soc. London SerA, 317:415 (1986)), PCR mutagenesis, or other known techniques can be performed on the cloned DNA to produce the peptide or composition of the invention.
Another subject of the invention relates to a protective peptide consisting of the amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 55 to 57, 59, 60, 68, 69, 71, 72, 79 to 84, 91 to 132, 175 to 179, 185 to 203, 223 to 234, and 235 to 425 which have been shown to provide protection against S. agalactiae (see Examples).
Another subject of the invention relates to one or more nucleic acid(s) encoding the at least two proteins comprised in the composition according to the invention and/or any of the protective peptides according to the invention.
Nucleic acid molecules of the present invention may be in the form of RNA, such as mRNA or cRNA, or in the form of DNA, including, for instance, cDNA and genomic DNA e.g. obtained by cloning or produced by chemical synthetic techniques or by a combination thereof. The DNA may be triple-stranded, double-stranded or single-stranded. Single-stranded DNA may be the coding strand, also known as the sense strand, or it may be the non-coding strand, also referred to as the anti-sense strand. Nucleic acid molecule as used herein also refers to, among others, single- and double-stranded DNA, DNA that is a mixture of single- and double-stranded RNA, and RNA that is a mixture of single- and double-stranded regions, hybrid molecules comprising DNA and RNA that may be single-stranded or, more typically, double-stranded, or triple-stranded, or a mixture of single- and double-stranded regions. In addition, nucleic acid molecule as used herein refers to triple-stranded regions comprising RNA or DNA or both RNA and DNA.
The nucleic acid may be a fragment of a nucleic acid occurring naturally in S. agalactiae. The nucleic acid also includes sequences that are a result of the degeneration of the genetic code. There are 20 natural amino acids, most of which are specified by more than one codon. Therefore, all nucleotide sequences are included in the invention which result in the peptide as defined above.
Preferred examples of the nucleic acid(s) encoding the at least two proteins comprised in the composition according to the invention and/or any of the protective peptides according to the invention are those comprising or consisting of at least one nucleic acid sequence selected from the group consisting of SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 61 to 66, SEQ ID NO: 73 to 78, SEQ ID NO: 85 to 90, SEQ ID NO: 133 to 174, SEQ ID NO: 180 to 184 and SEQ ID NO: 204 to 222. The above sequences are indicated in the Examples, Tables 7 to 12 and the attached Sequence listing.
Additionally, the nucleic acid may contain one or more modified bases. Such nucleic acids may also contain modifications e.g. in the ribose-phosphate backbone to increase stability and half life of such molecules in physiological environments. Thus, DNAs or RNAs with backbones modified for stability or for other reasons are “nucleic acid molecules” as that feature is intended herein. Moreover, DNAs or RNAs comprising unusual bases, such as inosine, or modified bases, such as tritylated bases, to name just two examples, are nucleic acid molecules within the context of the present invention. It will be appreciated that a great variety of modifications have been made to DNA and RNA that serve many useful purposes known to those of skill in the art. The term nucleic acid molecule as it is employed herein embraces such chemically, enzymatically or metabolically modified forms of nucleic acid molecule, as well as the chemical forms of DNA and RNA characteristic of viruses and cells, including simple and complex cells, inter alia. For example, nucleotide substitutions can be made which do not affect the peptide or protein or composition of the invention encoded by the nucleic acid, and thus any nucleic acid molecule which encodes an antigenic peptide or functionally active variant thereof or a composition of the invention as defined above is encompassed by the present invention.
Furthermore, any of the nucleic acid molecules encoding a peptide or composition of the invention can be functionally linked, using standard techniques such as standard cloning techniques, to any desired regulatory sequences, whether a S. agalactiae regulatory sequence or a heterologous regulatory sequence, heterologous leader sequence, heterologous marker sequence or a heterologous coding sequence to create a fusion protein.
The nucleic acid of the invention may be originally formed in vitro or in a cell in culture, in general, by the manipulation of nucleic acids by endonucleases and/or exonucleases and/or polymerases and/or ligases and/or recombinases or other methods known to the skilled practitioner to produce the nucleic acids.
In one embodiment of the invention, the nucleic acid(s) according to the invention is/are located in a vector or a cell other than S. agalactiae. 
A vector may further include nucleic acid sequences that permit it to replicate in the host cell, such as an origin of replication, one or more therapeutic genes and/or selectable marker genes and other genetic elements known in the art such as regulatory elements directing transcription, translation and/or secretion of the encoded peptide or protein. The vector may be used to transduce, transform or infect a cell, thereby causing the cell to express nucleic acids and/or proteins other than those native to the cell. The vector optionally includes materials to aid in achieving entry of the nucleic acid into the cell, such as a viral particle, liposome, protein coating or the like. Numerous types of appropriate expression vectors for protein expression are known in the art, which may be used in standard molecular biology techniques. Such vectors are selected from among conventional vector types including insects, e.g., baculovirus expression, or yeast, fungal, bacterial or viral expression systems. Other appropriate expression vectors, of which numerous types are known in the art, can also be used for this purpose. Methods for obtaining such expression vectors are well-known (see, e.g. Sambrook et al., Molecular Cloning. A Laboratory Manual, 2nd edition, Cold Spring Harbor Laboratory, New York (1989)). In one embodiment, the vector is a viral vector. Viral vectors include, but are not limited to, retroviral and adenoviral vectors.
Suitable host cells or cell lines for transfection by this method include bacterial cells. For example, the various strains of E. coli are well-known as host cells in the field of biotechnology. Various strains of B. subtilis, Pseudomonas, Streptomyces, and other bacilli and the like may also be employed in this method. Many strains of yeast cells known to those skilled in the art are also available as host cells for expression of the peptides of the present invention. Other fungal cells or insect cells such as Spodoptera frugipedera (Sf9) cells may also be employed as expression systems. Alternatively, mammalian cells, such as human 293 cells, Chinese hamster ovary cells (CHO), the monkey COS-1 cell line or murine 3T3 cells derived from Swiss, BALB/c or NIH mice may be used. Still other suitable host cells, as well as methods for transfection, culture, amplification, screening, production, and purification are known in the art.
A further subject of the invention relates to a method of producing the composition according to the invention or the protective peptide according to the invention, comprising

    (a) introducing the one or more nucleic acids into a host cell;
    (b) expressing the protein(s) and/or peptide(s) encoded by the nucleic acid by culturing the host cell under conditions conducive to the expression of the protein(s) and/or peptide(s); and
    (c) collecting and/or isolating the expressed protein(s) and/or peptide(s) of step (b).


A peptide or composition of the invention or component thereof may be produced by expressing a nucleic acid of the invention in a suitable host cell. The nucleic acid encoding the peptide/protein can be introduced into a host cell by any conventional technique. The host cells can e.g. be transfected, e.g. by conventional means such as electroporation with at least one expression vector containing a nucleic acid of the invention under the control of a transcriptional regulatory sequence. The transfected or transformed host cell is then cultured under conditions that allow expression of the protein. The expressed protein is recovered, isolated, and optionally purified from the cell (or from the culture medium, if expressed extracellularly) by appropriate means known to one of skill in the art. For example, the proteins are isolated in soluble form following cell lysis, or extracted using known techniques, e.g. in guanidine chloride. If desired, the peptides or fragments of the invention are produced as a fusion protein. Such fusion proteins are those described above. Alternatively, for example, it may be desirable to produce fusion proteins to enhance expression of the protein in a selected host cell or to improve purification. The molecules comprising the peptides and compositions of this invention may be further purified using any of a variety of conventional methods including, but not limited to: liquid chromatography such as normal or reversed phase, using HPLC, FPLC and the like; affinity chromatography (such as with inorganic ligands or monoclonal antibodies); size exclusion chromatography; immobilized metal chelate chromatography; gel electrophoresis; and the like. One of skill in the art may select the most appropriate isolation and purification techniques without departing from the scope of this invention. Such purification provides the peptide/protein/composition in a form substantially free from other proteinaceous and non-proteinaceous materials of the microorganism.
Still another subject of the invention relates to a pharmaceutical composition, especially a vaccine, comprising

    (i) the composition according to the invention and/or at least one protective peptide according to the invention; and
    (ii) optionally a pharmaceutically acceptable carrier or excipient.


A peptide or composition of the invention may be used for methods for immunizing or treating humans and/or animals with the disease caused by infection with S. agalactiae. Therefore, the peptide or composition may be used within a pharmaceutical composition. The pharmaceutical composition of the present invention may further encompass pharmaceutically acceptable carriers and/or excipients. The pharmaceutically acceptable carriers and/or excipients useful in this invention are conventional and may include buffers, stabilizers, diluents, preservatives, and solubilizers. Remington's Pharmaceutical Sciences, by E. W. Martin, Mack Publishing Co., Easton, Pa., 15th Edition (1975), describes compositions and formulations suitable for pharmaceutical delivery of the (poly)peptides/proteins herein disclosed.
If the pharmaceutical composition comprises at least two protective proteins as defined above, the proteins of subgroup i) to vi) may be formulated into one or more pharmaceutical composition(s). Additionally, the two or more pharmaceutical compositions may be administered together, simultaneously or consecutively.
In general, the nature of the carrier or excipients will depend on the particular mode of administration being employed. For instance, parenteral formulations usually comprise injectable fluids that include pharmaceutically and physiologically acceptable fluids such as water, physiological saline, balanced salt solutions, aqueous dextrose, glycerol or the like as a vehicle. For solid compositions (e.g. powder, pill, tablet, or capsule forms), conventional non-toxic solid carriers can include, for example, pharmaceutical grades of mannitol, lactose, starch, or magnesium stearate. In addition to biologically neutral carriers, pharmaceutical compositions to be administered can contain minor amounts of non-toxic auxiliary substances, such as wetting or emulsifying agents, preservatives, and pH buffering agents and the like, for example sodium acetate or sorbitan monolaurate.
In a preferred embodiment the pharmaceutical composition further comprises an immunostimulatory substance such as an adjuvant. The adjuvant can be selected based on the method of administration and may include mineral oil-based adjuvants such as Freund's complete and incomplete adjuvant, Montanide incomplete Seppic adjuvant such as ISA or ISA206 (SEPPIC, Paris, France), oil in water emulsion adjuvants such as the Ribi adjuvant system, syntax adjuvant formulation containing muramyl dipeptide, IC31® (Intercell; a synthetic adjuvant comprising the peptide motif KLK [WO 02/32451] and an oligonucleotide [WO 01/93905]), or aluminum salt adjuvants, preferably aluminum hydroxide or aluminum phosphate.
In a more preferred embodiment the immunostimulatory substance is selected from the group comprising polycationic polymers, especially polycationic peptides such as polyarginine, immunostimulatory deoxynucleotides (ODNs), especially Oligo(dIdC)13, peptides containing at least two LysLeuLys motifs, especially KLKLLLLLKLK, neuroactive compounds, especially human growth hormone, alum, adjuvants and combinations thereof. Preferably the combination is either a polycationic polymer and immunostimulatory deoxynucleotides or of a peptide containing at least two LysLeuLys motifs and immunostimulatory deoxynucleotides. In a still more preferred embodiment the polycationic polymer is a polycationic peptide.
The term “Oligo(dIdC)13” as used in the present invention means a phosphodiester backboned single-stranded DNA molecule containing 13 deoxy (inosine-cytosine) motifs, also defined by the term [oligo-d(IC)13]. The exact sequence is 5′-dIdCdIdCdIdCdIdCdIdCdIdCdIdCdIdCdIdCdIdCdIdCdIdCdIdC-3′. Oligo(dIdC)13 can also be defined by the terms (oligo-dIC26); oligo-dIC26-mer; oligo-deoxy IC, 26-mer; or oligo-dIC, 26-mer, as specified for example in WO 01/93903 and WO 01/93905.
In an even more preferred embodiment of the invention the immunostimulatory substance is at least one immunostimulatory nucleic acid. Immunostimulatory nucleic acids are e.g. natural or artificial CpG containing nucleic acids, short stretches of nucleic acids derived from non-vertebrates or in form of short oligonucleotides (ODNs) containing non-methylated cytosine-guanine dinucleotides (CpG) in a defined base context (e.g. as described in WO 96/02555). Alternatively, also nucleic acids based on inosine and cytidine as e.g. described in WO 01/93903, or deoxynucleic acids containing deoxy-inosine and/or deoxyuridine residues (described in WO 01/93905 and WO 02/095027) may preferably be used as immunostimulatory nucleic acids in the present invention. Preferably, mixtures of different immunostimulatory nucleic acids are used in the present invention. Additionally, the aforementioned polycationic compounds may be combined with any of the immunostimulatory nucleic acids as aforementioned. Preferably, such combinations are according to the ones described in WO 01/93905, WO 02/32451, WO 01/54720, WO 01/93903, WO 02/13857, WO 02/095027 and WO 03/047602.
In addition or alternatively, such a vaccine composition may comprise a neuroactive compound. Preferably, the neuroactive compound is human growth factor, e.g. described in WO 01/24822. Also preferably, the neuroactive compound is combined with any of the polycationic compounds and/or immunostimulatory nucleic acids as defined above.
In a highly preferred embodiment of the invention, the adjuvants are those used in the Examples, e.g. Complete Freund's adjuvant, aluminum hydroxide or/and an adjuvant comprising the KLKLLLLLKLK peptide and [dIdC]13 phosphodiester ssDNA, such as IC31® (Intercell AG, Vienna, Austria; described above).
The composition may be used e.g. for immunization or treatment of a subject. The pharmaceutical composition encompasses at least one peptide or composition of the invention; however, it may also contain a cocktail (i.e., a simple mixture) containing different peptides and/or compositions of the invention, optionally mixed with different antigenic peptides or proteins of other pathogens. Such mixtures of these peptides, polypeptides, proteins or fragments or variants thereof are useful e.g. in the generation of desired antibodies to a wide spectrum of S. agalactiae isolates. The (poly)peptide(s)/composition(s) of the present invention may also be used in the form of a pharmaceutically acceptable salt. Suitable acids and bases which are capable of forming salts with the peptides of the present invention are well known to those of skill in the art, and include inorganic and organic acids and bases.
Still another subject of the invention relates to a pharmaceutical composition comprising

    (i) the one or more nucleic acid(s) according to the invention or one or more nucleic acid(s) complementary thereto, and
    (ii) optionally a pharmaceutically acceptable carrier or excipient.


The nucleic acid sequences, alone or in combination with other nucleic acid sequences encoding peptides/proteins/compositions or antibodies or directed to other pathogenic microorganisms, may further be used as components of a pharmaceutical composition. The composition may be used for immunizing or treating humans and/or animals with the disease caused by infection with S. agalactiae. 
The pharmaceutically acceptable carrier or excipient may be as defined above.
In another embodiment, the nucleic acid sequences of this invention, alone or in combination with nucleic acid sequences encoding other antigens or antibodies from other pathogenic microorganisms, may further be used in compositions directed to actively induce a protective immune response in a subject to the pathogen. These components of the present invention are useful in methods for inducing a protective immune response in humans and/or animals against infection with S. agalactiae. 
For use in the preparation of the therapeutic or vaccine compositions, nucleic acid delivery compositions and methods are useful, which are known to those of skill in the art. The nucleic acids of the present invention or one or more nucleic acid(s) complementary thereto may be employed in the methods of this invention or in the compositions described herein as DNA sequences, either administered as naked DNA, or associated with a pharmaceutically acceptable carrier and provide for in vivo expression of the antigen, peptide or polypeptide. So-called “naked DNA” may be used to express the peptide or composition of the invention in vivo in a patient. (See, e.g., J. Cohen, Science, 259:1691-1692, which describes similar uses of “naked DNA”). For example, “naked DNA” associated with regulatory sequences may be administered therapeutically or as part of the vaccine composition e.g., by injection.
Alternatively, a nucleic acid encoding a peptide or composition of the invention or a nucleic acid complementary thereto may be used within a pharmaceutical composition, e.g. in order to express the peptide or composition of the invention in vivo, e.g., to induce antibodies.
A preferred embodiment of the invention relates to a pharmaceutical composition, wherein the nucleic acid is comprised in a vector and/or a cell other than S. agalactiae. Vectors and cells suitable in the context of the present invention are described above. Vectors are particularly employed for a DNA vaccine. An appropriate vector for delivery may be readily selected by one of skill in the art. Exemplary vectors for in vivo gene delivery are readily available from a variety of academic and commercial sources, and include, e.g., adeno-associated virus (International patent application No. PCT/US91/03440), adenovirus vectors (M. Kay et al., Proc. Natl. Acad. Sci. USA, 91:2353 (1994); S. Ishibashi et al., J. Clin. Invest., 92:883 (1993)), or other viral vectors, e.g., various poxviruses, vaccinia, etc. Recombinant viral vectors, such as retroviruses or adenoviruses, are preferred for integrating the exogenous DNA into the chromosome of the cell.
Another subject of the invention relates to a method for producing antibodies, characterized by the following steps:

    (a) administering an effective amount of the composition according to the invention and/or at least one protective peptide according to the invention to an animal; and
    (b) isolating the antibodies produced by the animal in response to the administration of step (a) from the animal.


A further subject of the invention relates to a method for producing antibodies, characterized by the following steps:

    (a) contacting a B cell with an effective amount of the composition according to the invention and/or at least one protective peptide according to the invention;
    (b) fusing the B cell of step (a) with a myeloma cell to obtain a hybridoma cell; and
    (c) isolating the antibodies produced by the cultivated hybridoma cell.


Also included in the scope of the invention is the production of antibodies against a peptide or composition according to the invention. This includes, for example, monoclonal and polyclonal antibodies, chimeric, single chain, and humanized antibodies, as well as Fab fragments, or the product of a Fab expression library, which are able to specifically bind to the peptide or composition according to the invention.
In a preferred embodiment the antibody is a monoclonal, polyclonal, chimeric or humanized antibody or functionally active fragment thereof. In another preferred embodiment the functionally active fragment comprises a Fab fragment.
Antibodies generated against the peptide or composition according to the invention can be obtained by direct injection of the peptide or composition according to the invention into an animal or administering of the peptide or composition according to the invention to an animal, preferably a non-human. The antibody so obtained will then bind the peptide or composition according to the invention. Such antibodies can then be used to isolate reactive antigens, peptide or proteins from a tissue expressing those.
For preparation of monoclonal antibodies, any technique known in the art, which provides antibodies produced by continuous cell line cultures, e.g. a hybridoma cell line, can be used.
Techniques described for the production of single chain antibodies (U.S. Pat. No. 4,946,778) can be adapted to produce single chain antibodies to the antigenic peptides or compositions according to the invention. Also, transgenic mice or other organisms such as other mammals may be used to express humanized antibodies to the antigenic peptides or compositions according to the invention.
Antibodies may be also produced using a hybridoma cell line. Hybridoma cell lines expressing desirable monoclonal antibodies are generated by well-known conventional techniques. The hybridoma cell can be generated by fusing a normal-activated, antibody-producing B cell with a myeloma cell. In the context of the present invention the hybridoma cell is able to produce an antibody specifically binding to the antigenic peptide or composition according to the invention.
Similarly, desirable high titer antibodies are generated by applying known recombinant techniques to the monoclonal or polyclonal antibodies developed to these peptides/proteins/compositions (see, e.g., PCT Patent Application No. PCT/GB85/00392; British Patent Application Publication No. GB2188638A; Amit et al., Science, 233:747-753 (1986); Queen et al., Proc. Natl. Acad. Sci. USA, 86:10029-10033 (1989); PCT Patent Application No. WO90/07861; Riechmann et al., Nature, 332:323-327 (1988); Huse et al., Science, 246:1275-1281 (1988)).
Particularly, the antibody may be produced by initiating an immune response in a non-human animal by administrating a peptide or composition of the invention to an animal, removing an antibody-containing body fluid from said animal, and producing the antibodies by subjecting said antibodies containing body fluid to further purification steps.
Alternatively, the antibody may be produced by initiating an immune response in a non-human animal by administrating a peptide or composition, as defined in the present invention, to said animal, removing the spleen or spleen cells from said animal and/or producing hybridoma cells of said spleen or spleen cells, selecting and cloning hybridoma cells specific for the peptide or composition according to the invention and producing the antibody by cultivation of said cloned hybridoma cells.
Alternatively, the antibody may be produced employing a phage display antibody library. The method is based on the selective binding of one or more members of a phage display antibody library to a surface-bound antigen. The method may e.g. be carried out as follows: an antigen of choice is immobilized to a solid surface, such as nitrocellulose, magnetic beads, a column matrix or, the most widely used, plastic surfaces as polystyrole tubes or 96-well plates. The antibody phages are incubated with the surface-bound antigen, followed by thorough washing to remove the excess nonbinders. The bound antibody phage can subsequently be eluted and e.g. amplified by infection of Escherichia coli. This method allows the detection of a single antibody phage and as it can be selected by e.g. its resistance marker, it can give rise to a bacterial colony after elution. The isolation of antibodies using phage display antibody libraries has been described in more details by Mancini et al., New Microbiol. 2004 October; 27(4):315-328 and Pini et al., Curr Protein Pept Sci. 2004 December; 5(6):487-496.
In a preferred embodiment the antibodies produced according to a method of the invention are additionally purified. Methods of purification are known to the skilled artisan.
The antibody may be used in methods for treating an infection. Accordingly, still another subject of the invention relates to a pharmaceutical composition, especially a vaccine, comprising the antibody produced according to the invention. The pharmaceutical composition may encompass further components as detailed above. The composition may further encompass substances increasing their capacity to stimulate T cells. These include T helper cell epitopes, lipids or liposomes or preferred modifications as described in WO01/78767. Another way to increase the T cell stimulating capacity of epitopes is their formulation with immune stimulating substances for instance cytokines or chemokines like interleukin-2, -7, -12, -18, class I and II interferons (IFN), especially IFN-gamma, GM-CSF, TNF-alpha, flt3-ligand and others.
Another subject of the invention relates to a mixture of antibodies against the at least two proteins of the composition according to the invention and/or against the at least one protective peptide according to the invention. The mixture of antibodies may be further characterized and produced as described above.
Methods of producing antibodies, mixtures of antibodies, as well as the use of antibodies are also described in Examples 4 and 5, and FIGS. 4, 8, and 10 to 13.
Another subject of the invention relates to the use of the composition according to the invention and/or at least one protective peptide according to the invention and/or one or more of the nucleic acid(s) according to the invention for the manufacture of a medicament for the immunization or treatment of a subject, preferably against S. agalactiae, more preferably against pneumonia, septicemia, meningitis, fever, vomiting, poor feeding, irritability, urinary tract infection and/or vaginal infection caused by S. agalactiae. 
The peptides, proteins, compositions or the nucleic acids of the invention are generally useful for inducing an immune response in a subject. The vaccine used for immunization may be administered to a subject susceptible to infection by S. agalactiae, preferably mammals, and still more preferably humans, in any conventional manner, including oral, topical, intranasal, intramuscular, intra-lymph node, intradermal, intraperitoneal, subcutaneous, and combinations thereof, but most preferably through intramuscular injection. The volume of the dose for intramuscular administration is preferably up to about 5 ml, still more preferably between 0.5 ml and 3 ml, and most preferably about 1 to 2 ml. The volume of the dose when subcutaneous injection is the selected administration route is preferably up to about 5 ml, still more preferably between 0.5 ml and 3 ml, and most preferably about 1 to 2 ml. The amount of substance in each dose should be enough to confer effective immunity against and decrease the risk of developing clinical signs resulting from S. agalactiae infection to a subject receiving a vaccination therewith. Preferably, the unit dose of protein should be up to about 5 μg protein/kg body weight, more preferably between about 0.2 to 3 μg, still more preferably between about 0.3 to 1.5 μg, more preferably between about 0.4 to 0.8 μg, and still more preferably about 0.6 μg. Alternative preferred unit doses of protein could be up to about 6 μg protein/kg body weight, more preferably between about 0.05 to 5 μg, still more preferably between about 0.1 to 4 μg. The dose is preferably administered 1 to 3 times, e.g. with an interval of 1 to 4 weeks. Preferred amounts of protein per dose are from approximately 1 μg to approximately 1 mg, more preferably from approximately 5 μg to approximately 500 μg, still more preferably from approximately 10 μg to approximately 250 μg and most preferably from approximately 25 μg to approximately 100 μg.
In still another aspect of the invention the mixture of antibodies or the antibody produced according to the invention or functional fragment thereof is used for the manufacture of a medicament for the treatment of an infection, preferably a S. agalactiae infection. The treatment involves administering an effective amount of the antibody to a subject, preferably a mammal, more preferably a human. Thus, antibodies against the peptides or the composition of the present invention may be employed to inhibit and/or treat infections, particularly bacterial infections and especially infections arising from S. agalactiae. 
An “effective amount” of peptides, proteins, compositions or the nucleic acids of the invention or an antibody produced according to the invention may be calculated as that amount capable of exhibiting an in vivo effect, e.g. preventing or ameliorating a sign or symptom of infection, particularly S. agalactiae infection. Such amounts may be determined by one of skill in the art. Such a substance may be administered in any conventional manner, including oral, topical, intranasal, intramuscular, intra-lymph node, intradermal, intraperitoneal, subcutaneous, and combinations thereof, but preferably intramuscularly or subcutaneously. However, it may also be formulated to be administered by any other suitable route, including orally or topically. The selection of the route of delivery and dosage of such therapeutic compositions is within the skill of the art.
Treatment in the context of the present invention refers to both therapeutic treatment and prophylactic or preventive measures, wherein the object is to prevent or slow down (lessen) the targeted pathologic condition or disorder. Those in need of treatment include those already with the disorder as well as those prone to have the disorder or those in whom the disorder is to be prevented.
Another subject of the invention relates to a method of diagnosing a S. agalactiae infection comprising the steps of:

    (a) contacting a sample obtained from a subject with the composition according to the invention and/or at least one protective peptide according to the invention; and
    (b) detecting the presence of an antibody against the protective peptide, the functionally active variant and/or the composition in the sample,

wherein the presence of the antibody is indicative for the S. agalactiae infection.



Another subject of the invention relates to a method of diagnosing a S. agalactiae infection comprising the steps of:

    (a) contacting a sample obtained from a subject with the mixture of antibodies according to the invention; and
    (b) detecting the presence of the at least two proteins of the composition according to the invention and/or of the at least one protective peptide according to the invention in the sample,

wherein the presence of the at least two proteins and/or of the at least one protective peptide is indicative for the S. agalactiae infection.



The protective peptides or compositions of the invention or alternatively a mixture of antibodies may be used for the detection of S. agalactiae. Preferably such detection is for diagnosis, more preferably for the diagnosis of a disease, most preferably for the diagnosis of a S. agalactiae infection. The protective peptides or compositions may be used to detect the presence of a S. agalactiae-specific antibody or fragment thereof e.g. in a sample obtained from a subject. Alternatively, the mixture of antibodies may be used to detect the presence of S. agalactiae proteins, e.g. in a sample obtained from a subject. The sample may be e.g. a blood sample.
The present invention also relates to diagnostic assays such as quantitative and diagnostic assays for detecting levels of the proteins, compositions and/or mixtures of antibodies of the present invention in cells and tissues or body fluids, including determination of normal and abnormal levels. Assay techniques that can be used to determine levels of a peptide, a composition or an antibody, in a sample derived from a host are well known to those of skill in the art. Such assay methods include radioimmunoassays, competitive-binding assays, Western Blot analysis and ELISAs. Among these, ELISAs frequently are preferred. An ELISA initially comprises preparing an antibody or antibodies specific to the peptide or composition, preferably a monoclonal antibody. In addition, a reporter antibody generally is prepared which binds to the monoclonal antibody. The reporter antibody is attached to a detectable reagent such as radioactive, fluorescent or enzymatic reagent, such as horseradish peroxidase enzyme.
The peptides or compositions of the present invention may also be used for the purpose of or in connection with an array. More particularly, at least one of the peptides or compositions of the present invention may be immobilized on a support. Said support typically comprises a variety of peptides/proteins whereby the variety may be created by using one or several of the peptides or compositions of the present invention. The characterizing feature of such array as well as of any array in general is the fact that at a distinct or predefined region or position on said support or a surface thereof, a distinct polypeptide is immobilized. Because of this any activity at a distinct position or region of an array can be correlated with a specific polypeptide. The number of different peptides or antibodies of the present invention immobilized on a support may range from as little as 10 to several 1000 different peptides or compositions of the present invention. Alternatively, antibodies produced according to the present invention may be used to detect peptides or compositions of the invention.
The manufacture of such arrays is known to the one skilled in the art and, for example, described in U.S. Pat. No. 5,744,309. The array preferably comprises a planar, porous or non-porous solid support having at least a first surface. Preferred support materials are, among others, glass or cellulose. It is also within the present invention that the array is used for any of the diagnostic applications described herein. Apart from the peptides or antibodies of the present invention also the nucleic acid molecules according to the present invention may be used for the generation of an array as described above.
Another subject of the invention relates to a method for diagnosing an infection with S. agalactiae comprising the steps of:

    (a) contacting a sample obtained from a subject with a primer and/or a probe specific for the one or more nucleic acid(s) according to the invention; and
    (b) detecting the presence of one or more nucleic acid(s) according to the invention in the sample,

wherein the presence of the one or more nucleic acid(s) is indicative for the S. agalactiae infection.



A series of methods for detecting nucleic acids in samples by using specific primers and/or probes is known in the art. In general, these methods are based on the specific binding of a primer or probe to the nucleic acid in question. The methods may involve amplification of the nucleic acid, e.g. RNA or DNA, before the actual detection step. Therefore, primers may be used to specifically induce transcription and/or amplification of RNA or DNA in order to generate a detectable amount of nucleic acid. Suitable well known techniques may be PCR and RT-PCR. Suitable primers and probes for the method of the invention may be produced based on sequence information provided in the present application. Guidelines and computer-assisted programs (e.g Primer Express®, Applied Biosystems, Foster City, Calif., USA) for designing primers and probes to a specific nucleic acid are known to the person skilled in the art.
After the amplification step the amplified nucleic acid, in general DNA, may be detected e.g. by its size (e.g. involving agarose gel electrophoresis) or using labeled probes which specifically bind to the amplified nucleic acid. The probes may be labeled with a dye, radioactive marker, a fluorescent marker, an enzyme-linked marker or any other marker.
For example, FRET (Forster resonance energy transfer) may be used for the detection of the nucleic acid of the invention. In FRET, a donor fluorophore molecule absorbs excitation energy and delivers this via dipole-dipole interaction to a nearby acceptor fluorophore molecule. This process only occurs when the donor and acceptor molecules are sufficiently close to one another. Several different strategies for determining the optimal physical arrangement of the donor and acceptor moieties are known to the skilled practitioner. For this, a fluorescent donor is excited at its specific fluorescence excitation wavelength. By a long-range dipole-dipole coupling mechanism, this excited state is then nonradiatively transferred to a second molecule, the acceptor. The donor returns to the electronic ground state. The described energy transfer mechanism is termed “Forster resonance energy transfer” (FRET). The process involves measuring fluorescence as FRET donor and acceptor moieties are brought together as a result of DNA hybridization. For examples two probes each labeled with a suitable marker hybridize to the nucleic acid of the invention within a distance which allows FRET to occur. Suitable markers include Cyan 500, Cy5, Cy3, SYBR Green I, fluorescein, HEX, Red 610 and Red 640, wherein the two marker involved have to be selected based on there excitation and emission spectrums as known by the skilled person. A suitable system for the detection of nucleic acids is the LightCycler® (Roche Diagnostics).
Another subject of the invention relates to a method for identifying a ligand capable of binding the composition according to the invention and/or at least one protective peptide according to the invention comprising:

    (a) providing a test system comprising the peptide and/or composition,
    (b) contacting the test system with a test compound, and
    (c) detecting a signal generated in response to the binding of the test compound to the peptide and/or composition.


More particularly, the method may be carried out by contacting an isolated or immobilized protective peptide or composition according to the invention with a candidate ligand under conditions to permit binding of the candidate ligand to the peptide, wherein the test system comprises a component capable of providing a detectable signal in response to the binding of the candidate ligand to said peptide; and detecting the presence or absence of a signal generated in response to the binding of the ligand to the peptide. The ligand may be an agonist or an antagonist.
Test systems for detection binding of a ligand are known to the skilled artisan and include e.g. binding assays with labeled ligand such as radioligands, fluorescence-labeled ligands or enzyme-labeled ligands.
The test compound can be any test compound either naturally occurring or chemically synthesized. Naturally occurring test compounds include in particular antibodies, preferably those showing similarity to the antibodies of the invention. In one preferred embodiment of the invention the test compound is provided in the form of a chemical compound library. Chemical compound libraries include a plurality of chemical compounds and have been assembled from any of multiple sources, including chemically synthesized molecules and natural products, or have been generated by combinatorial chemistry techniques. They are especially suitable for high throughput screening. They may be comprised of chemical compounds of a particular structure or compounds of a particular creature such as a plant.
A further subject of the invention relates to the use of the composition according to the invention and/or at least one protective peptide according to the invention for the isolation and/or purification and/or identification of an interaction partner of the composition and/or peptide. The isolation and/or purification and/or identification of the ligand may be carried out as detailed above or as known to the person skilled in the art. In a preferred embodiment of the invention an affinity device may be used. The affinity device may comprise at least a support material and any antigenic peptide or composition according to the present invention, which is attached to the support material. Because of the specificity of the protective peptides and/or compositions according to the present invention for their target cells or target molecules or their interaction partners, the peptides and/or compositions allow a selective removal of their interaction partner(s) from any kind of sample applied to the support material provided that the conditions for binding are met. The sample may be a biological or medical sample, including but not limited to, fermentation broth, cell debris, cell preparation, tissue preparation, organ preparation, blood, urine, lymph liquid, liquor and the like. The peptide or composition may be attached to the matrix in a covalent or non-covalent manner. Suitable support material is known to the one skilled in the art and can be selected from the group comprising cellulose, silicon, glass, aluminium, paramagnetic beads, starch and dextrane.
The present invention is further illustrated by the following Figures, Examples and the Sequence listing, from which further features, embodiments and advantages may be taken. It is to be understood that the present examples are given by way of illustration only and not by way of limitation of the disclosure.



BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 shows the protection achieved by passive immunization with selected hyper-immune rabbit sera generated by immunization of rabbits with S. agalactiae antigens in a mouse lethality model. CD-1 mice (10 mice per group) were immunized intraperitoneally with 150 μl hyper-immune rabbit sera 1-3 hours before they were intraperitoneally challenged. (A) gbs1309p, gbs1478p, gbs2018p, Sip and PBS-induced hyperimmune sera, challenge with 1×107 cfu C388/90 (serotype Ia/c). (B) gbs1478p, gbs2018p, Sip and PBS-induced hyperimmune sera, challenge with 5×106 cfu ATCC12401 (serotype Ib). (C) gbs0233p, gbs1087p, gbs1477p, Sip and PBS-induced hyperimmune sera, challenge with 1×108 cfu ATCC12403 (serotype III). (D) gbs0233p, gbs1087p, gbs2018p, Sip and PBS-induced hyperimmune sera, challenge with 1×108 cfu ATCC49447 (serotype V). Survival was monitored for 14 days post-challenge. Numbers of surviving mice are plotted as percentage of total mice.
FIG. 2 shows the protection achieved with a combination of S. agalactiae antigen-specific hyperimmune rabbit sera in a mouse lethality model. CD-1 mice (10 mice per group) were immunized intraperitoneally with different combinations of hyperimmune rabbit sera (150 μl per hyper-immune rabbit sera) 1-3 hours before an intraperitoneal challenge. Sip- and PBS-induced sera were used as positive and negative controls, respectively. Mice were immunized with combinations of sera induced by gbs 1087p, gbs1477p, gbs1478p and gbs2018p; challenge with (A) 5×106 cfu ATCC12401 (serotype Ib); (B) 1×108 cfu ATCC12403 (serotype III) and (C) 1×108 cfu ATCC49447 (serotype V). Survival was monitored for 11 days post-challenge. Numbers of surviving mice are plotted as percentage of total mice.
FIG. 3 shows the surface staining of the serotype III GBS strain ATCC12403. The results for the hyperimmune rabbit sera (black) are shown in comparison to those for the respective preimmune sera (white).
FIG. 4 shows an opsonophagocytic killing assay with hyperimmune rabbit sera and different GBS strains. (A) Serotype II GBS strain AC3912, not suitable for animal testing. (B) Serotype V GBS strain BAA23, used for animal testing. PI, preimmune sera; HI, hyperimmune sera. GBS cells in the exponential phase were opsonised with 200-fold diluted sera in the presence of 5% guinea pig complement for 60 minutes. Phagocytic cells (RAW264.7) were added to opsonised bacteria and incubated for an additional 60 minutes at 37° C. Surviving bacteria were counted on agar plates after overnight incubation at 37° C. Percentage of killing was calculated based on CFU obtained after incubation with the different hyperimmune sera relative to CFU obtained with preimmune sera at 0 min and after 60 min of incubation as described under experimental procedure.
FIG. 5 A-D show the sequence alignment of protein sequences homologous to gbs1477 from genomic and sequenced strains. Alignment of sequences was performed using the software from Vector NTI (Suite 7.1; Invitrogen, Austria). The name on the left of the sequence indicates the strain name Amino acids in bold, residue identical in at least 50% of sequences. *, indicates position of STOP codon.
FIG. 6 shows active protection of adult mice by immunizing with a combination of GBS antigens. CD-1 mice (10 mice per group) were immunized with different combinations of the recombinant proteins (gbs1087p, gbs1477p, gbs1478p and gbs2018p; 25 μg each). As positive control 25 μg Sip protein (open circle) was used. For the negative control, PBS (open square) was used with ALUM 1%. One week after the last booster immunization, mice were challenged with (A) 3.5×106 cfu 12401; (B) 8.8×107 cfu 12403 or with (C) 1.1×108 cfu 49447. Numbers of surviving mice are plotted as a percentage of the total number of mice.
FIG. 7 shows active protection of adult mice by immunizing with single GBS antigens. CD-1 mice (10 mice per group) were immunized with different recombinant proteins (gbs0233p, gbs1087p, gbs1309p, gbs1477p, gbs1478p and gbs2018p; 50 μg each). As positive control 50 μg Sip protein (open circle) was used. For the negative control, PBS (open square) was used with ALUM 1%. One week after the last booster immunization, mice were challenged with (A) 3.5×106 cfu 12401; (B) 8.8×107 cfu 12403 or with (C) 1.1×108 cfu 49447. Numbers of surviving mice are plotted as a percentage of the total number of mice.
FIG. 8 shows the protection of neonatal mice by immunization of mothers passively with rabbit sera generated with a combination of recombinant GBS antigens. Pregnant CD-1 mice were immunized at day 18 post gestation with 500 μl of combinations of gbs1087p, gbs1477p, gbs1478p and gbs2018p-induced sera or PBS-induced control sera. Neonates were challenged within 24-38 hours after birth with lethal challenge doses of (A) 1.2×107 cfu C388/90; (B) 1.3×106 cfu 12403; (C) 5.7×106 cfu BAA23 or (D) 1.8×108 cfu 2603V/R. Numbers of surviving neonates are plotted as a percentage of the total number of challenged neonates.
FIG. 9 shows active protection of adult mice by immunizing with different clade gbs1477p proteins. CD-1 mice (10 mice per group) were immunized with different clade proteins of gbs1477p (gbs1477p-2603V/R, gbs1477p-49447, gbs1477p-C388/90, gbs1477p-126H4A and gbs1477p-NEM316; 50 μg each). As negative control, PBS (open square) was used with ALUM 1%. One week after the last booster immunization, mice were challenged with (A) 1.4×107 cfu C388/90; (B) 1.2×108 cfu 12403 or with (C) 1.6×108 cfu BAA23. Numbers of surviving mice are plotted as a percentage of the total number of mice.
FIG. 10 shows the protection of adult mice by immunization with mouse mAbs against different recombinant GBS antigens. CD-1 mice were immunized intraperitoneally with 50 μg of the respective mouse mAb. 1 to 3 hours later, mice were challenged intraperitoneally with (A) 1.2×108 cfu 12403; (B) 1.5×108 cfu 12403 or (C) 1.1×108 cfu 49447. Numbers of surviving mice are plotted as a percentage of the total number of challenged mice.
FIG. 11 shows the protection of adult mice by immunization with a combination of five mouse mAbs against different recombinant GBS antigens. CD-1 mice were immunized intraperitoneally with 25 μg of each mouse mAb. 1 to 3 hours later, mice were challenged intraperitoneally with (A) 1.2×107 cfu C388/90; (B) 9.6×107 cfu 12403 or (C) 1.7×108 cfu BAA23. Numbers of surviving mice are plotted as a percentage of the total number of challenged mice.
FIG. 12 shows surface staining of the serotype III GBS strain ATCC12403. The results for the monoclonal antibodies (black) are shown with the buffer control (white).
FIG. 13 shows opsonophagocytic killing assay with hyperimmune rabbit sera and different GBS strains. Rabbit sera were tested in the opsonophagocytic killing assay at a serum dilution of 1:1,000, % killing was calculated in relation to the respective pre-immune sera. The mouse monoclonal antibodies were tested with two different amounts added to the opsonophagocytic killing assay, 1.0 and 0.1 μg. The % killing for the monoclonal antibodies was calculated in relation to the complement control.



EXAMPLES
Example 1
Group B Streptococcal Antigens and Combinations Thereof Inducing Protective Immune Responses Against Lethal Sepsis in an i.p. Challenge Model
Experimental Procedures
Cloning and Expression of Recombinant Group B Streptococcal Proteins
Cloning of Genes:
The gene of interest was amplified from genomic DNA of S. agalactiae ATCC12403 (serotype III) by PCR using gene specific primers. Apart from the gene specific part, the primers had restriction sites that aided in a directional cloning of the amplified PCR product. The gene annealing (specific) part of the primer ranged between 15-30 bases in length. The PCR products obtained were digested with the appropriate restriction enzymes and cloned into the pET28b (+) vector (Novagen) for His-tagged proteins. Once the recombinant plasmid was confirmed to contain the gene of interest, E. coli BL21 Star® cells (Invitrogen) that served as expression host were transformed. Cloning of the gbs1087, gbs1477 and gbs1478 genes has been performed using genomic DNA from strain S. agalactiae 6313 (serotype III) in the vector pET28a (+). The origin of the gene and position within the full length gene of the selected antigens are listed in Table 1. The amino acid and nucleic acid sequences are as follows:
Amino Acid Sequences:








SEQ ID NO: 1



Construct 1: gbs0233p



LCLALLTISGCQLTDTKKPGHTTIKVAAQSSTESSIMANIVTELIHHELGYNTTLISNLGS


 


STVTHQALLRGDADIAATRYTGTDITGTLGLKAVKDTKEASKIVKTEFQKRYNQTWYPTYG


 


FSDTYAFMVTKEFARQNKITKISDLKKLSTTMKAGVDSSWMNREGDGYTDFAKTYGFEFSH


 


IYPMQIGLVYDAVESNKMQSVLGYSTDGRISSYDLEILRDDKKFFPPYEASMVVNNSIIKK


 


DPKLKKLLHRLDGKINLKTMQNLNYMVDDKLLEPSVVAKQFLEKNHYFRGD


 


SEQ ID NO: 2



Construct 2: gbs1087p



MDSVGNQSQGNVLERRQRDAENRSQGNVLERRQRDVENKSQGNVLERRQRDAENKSQGNVL


 


ERRQRDAENRSQGNVLERRQRDAENRSQGNVLERRQRDAENRSQGNVLERRQRDAENRSQG


 


NVLERRQRDAENRSQGNVLERRQRDVENKSQGNVLERRQRDAENKSQGNVLERRQRDAENR


 


SQGNVLERRQRDAENRSQGNVLERRQRDAENRSQGNVLERRQRDAENRSQGNVLERRQRDA


 


ENRSQGNVLERRQRDAENRSQGNVLERRQRDAENRSQGNVLERRQRDAENRSQGNVLERRQ


 


RDAENKSQVGQLIG


 


SEQ ID NO: 3



Construct 3: gbs1309p



SVTYSQSERTVVFSFGEITFSRSRWTNGFETRIPVDEWLGLEKYKRYSIEFLYHVAKLATM


 


MPYRQVCKVIDSTLQTIITKDCVLKAVKFVEKLLKEKERYRFYLEEPPERKKVKKLYVEGD


 


GVMIKSTDSREERRYLDLTHFVIHTGSKKVSTKRYELQDKHEILQLNYDKAKYNLLDYIYN


 


NYEVDDDTILITNSDMGKGYTSRVFKELGKALKVKKHEHFWDIYHVKEKLSSYLRKYPIEL


 


TDFALDAVKKYNSDKLELVFDTVESLICDELEDQEFQKFKKKVLNNFKYIKPAHLRNLSNR


 


GIGIMESQHRKITYRMKRRGMYWSKWGISTMANMIILERANGLRELFFGSWRKVYSEYKEG


 


SFSAGRLFKKTDELDKFSKPLLKNGRKWSITGIK


 


SEQ ID NO: 4



Construct 4: gbs1477p



DDVTTDTVTLHKIVMPQAAFDNFTEGTKGKNDSDYVGKQINDLKSYFGSTDAKEIKGAFFV


 


FKNETGTKFITENGKEVDTLEAKDAEGGAVLSGLTKDTGFAFNTAKLKGTYQIVELKEKSN


 


YDNNGSILADSKAVPVKITLPLVNNQGVVKDAHIYPKNTETKPQVDKNFADKDLDYTDNRK


 


DKGVVSATVGDKKEYIVGTKILKGSDYKKLVWTDSMTKGLTFNNNVKVTLDGKDFPVLNYK


 


LVTDDQGFRLALNATGLAAVAAAAKDKDVEIKITYSATVNGSTIVEVPETNDVKLDYGNNP


 


TEESEPQEGTPANQEIKVIKDWAVDGTITDVNVAVKAIFTLQEKQTDGTWVNVASHEATKP


 


SRFEHTFTGLDNTKTYRVVERVSGYTPEYVSFKNGVVTIKNNKNSNDPTPINPSEPKVVTY


 


GRKFVKTNQANTERLAGATFLVKKEGKYLARKAGAATAEAKAAVKTAKLALDEAVKAYNDL


 


TKEKQEGQEGKTALATVDQKQKAYNDAFVKANYSYEWVADKKADNVVKLISNAGGQFEITG


 


LDKGTYSLEETQAPAGYATLSGDVNFEVTATSYSKGATTDIAYDKGSVKKDAQQVQNKKVT


 


IPQTGGIGT


 


SEQ ID NO: 5



Construct 5: gbs1478p



ESTVPENGAKGKLVVKKTDDQNKPLSKATFVLKTTAHPESKIEKVTAELTGEATFDNLIPG


 


DYTLSEETAPEGYKKTNQTWQVKVESNGKTTIQNSGDKNSTIGQNHEELDKQYPPTGIYED


 


TKESYKLEHVKGSVPNGKSEAKAVNPYSSEGEHIREIPEGTLSKRISEVGDLAHNKYKIEL


 


TVSGKTIVKPVDKQKPLDVVFVLDNSNSMNNDGPNFQRHNKAKKAAEALGTAVKDILGANS


 


DNRVALVTYGSDIFDGRSVDVVKGFKEDDKYYGLQTKFTIQTENYSHKQLTNNAEEIIKRI


 


PTEAPRAKWGSTTNGLTPEQQKQYYLSKVGETFTMKAFMEADDILSQVDRNSQKIIVHITD


 


GVPTRSYAINNFKLGASYESQFEQMKKNGYLNKSNFLLTDKPEDIKGNGESYFLFPLDSYQ


 


TQIISGNLQKLHYLDLNLNYPKGTIYRNGPVREHGTPTKLYINSLKQKNYDIFNFGIDISA


 


FRQVYNEDYKKNQDGTFQKLKEEAFELSDGEITELMKSFSSKPEYYTPIVTSSDASNNEIL


 


SKIQQQFEKVLIKENSIVNGTIEDPMGDKINLQLGNGQTLQPSDYTLQGNDGSIMKDSIAT


 


GGPNNDGGILKGVKLEYIKNKLYVRGLNLGEGQKVTLTYDVKLDDSFISNKFYDTNGRTTL


 


NPKSEDPNTLRDFPIPKIRDVREYPTITIKNEKKLGEIEFTKVDKDNNKLLLKGATFELQE


 


FNEDYKLYLPIKNNNSKVVTGENGKISYKDLKDGKYQLIEAVSPKDYQKITNKPILTFEVV


 


KGSIQNIIAVNKQISEYHEEGDKHLITNTHIPPKGIIPMTGGK


 


SEQ ID NO: 6



Construct 6: gbs2018p



DTSSGISASIPHKKQVNLGAVTLKNLISKYRGNDKAIAILLSRVNDFNRASQDTLPQLINS


 


TEAEIRNILYQGQIGKQNKPSVTTHAKVSDQELGKQSRRSQDIIKSLGFLSSDQKDILVKS


 


ISSSKDSQLILKFVTQATQLNNAESTKAKQMAQNDVALIKNISPEVLEEYKEKIQRASTKS


 


QVDEFVAEAKKVVNSNKETLVNQANGKKQEIAKLENLSNDEMLRYNTAIDNVVKQYNEGKL


 


NITAAMNALNSIKQAAQEVAQKNLQKQYAKKIERISSKGLALSKKAKEIYEKHKSILPTPG


 


YYADSVGTYLNRFRDKQTFGNRSVWTGQSGLDEAKKMLDEVKKLLKELQDLTRGTKEDKKP


 


DVKPEAKPEAKPNIQVPKQAPTEAAKPALSPEALTRLTTWYNQAKDLLKDDQVKDKYVDIL


 


AVQKAVDQAYDHVEEGKFITTDQANQLANKLRDALQSLELKDKKVAKPEAKPEAKPEAKPE


 


AKPEAKPEAKPEAKPEAKPDVKPEAKPDVKPEAKPEAKPEAKSEAKPEAKLEAKPEAKPAT


 


KKSVNTSGNLAAKKAIENKKYSKKLPST






Nucleic Acid Sequences:









SEQ ID NO: 7



Construct 1: gbs0233p



CTTTGCTTAGCCCTCTTAACGATTTCTGGTTGTCAATTAACCGATACTAAAAAACCTGGTC


 


ATACCACAATTAAGGTTGCTGCCCAAAGTTCTACAGAGTCTAGTATCATGGCAAATATTGT


 


CACCGAATTAATTCATCACGAATTAGGATACAACACAACTTTAATAAGCAATCTTGGTTCC


 


TCTACGGTTACTCACCAAGCTTTGCTCCGTGGTGATGCTGACATTGCTGCCACACGTTATA


 


CAGGAACAGACATCACAGGAACTCTTGGCTTAAAAGCTGTTAAAGACACTAAAGAAGCTTC


 


TAAGATTGTAAAAACTGAATTCCAAAAACGCTACAATCAAACTTGGTATCCTACTTATGGT


 


TTTTCTGATACTTATGCATTCATGGTTACTAAAGAGTTTGCCAGACAGAATAAAATCACCA


 


AGATCTCTGATCTCAAAAAGTTATCAACAACTATGAAGGCAGGGGTTGATAGTTCATGGAT


 


GAATCGCGAGGGAGATGGATACACTGATTTCGCTAAAACATACGGTTTTGAATTTTCACAT


 


ATTTACCCTATGCAAATTGGCTTAGTCTATGATGCAGTTGAAAGTAACAAAATGCAATCTG


 


TATTAGGCTACTCCACTGACGGTCGTATTTCGAGCTATGATTTAGAAATTTTAAGGGATGA


 


TAAAAAATTCTTTCCTCCTTATGAAGCCTCTATGGTTGTCAACAATTCTATCATCAAAAAA


 


GATCCTAAACTAAAAAAATTACTCCATCGACTCGATGGTAAAATCAATTTAAAAACGATGC


 


AAAACCTTAATTATATGGTAGATGATAAACTTTTAGAACCTTCAGTTGTTGCCAAACAATT


 


TTTAGAAAAAAACCATTATTTTAGAGGAGAT


 


SEQ ID NO: 8



Construct 2: gbs1087p



ATGGATAGTGTTGGAAATCAAAGTCAGGGCAATGTTTTAGAGCGTCGTCAACGTGATGCAG


 


AAAACAGAAGCCAAGGCAATGTTCTAGAGCGTCGTCAACGCGATGTTGAGAATAAGAGCCA


 


AGGCAATGTTTTAGAGCGTCGTCAACGTGATGCGGAAAACAAGAGCCAAGGCAATGTTTTA


 


GAGCGTCGTCAACGTGATGCAGAAAACAGAAGCCAAGGCAATGTTCTAGAGCGTCGTCAAC


 


GTGATGCAGAAAACAGAAGCCAAGGCAATGTTCTAGAGCGTCGTCAACGCGATGCAGAAAA


 


CAGAAGCCAAGGTAATGTTCTAGAGCGTCGTCAACGTGATGCAGAAAACAGAAGCCAAGGT


 


AATGTTCTAGAGCGTCGTCAACGTGATGCAGAAAACAGAAGCCAAGGTAATGTTCTAGAGC


 


GTCGTCAACGCGATGTTGAGAATAAGAGCCAAGGCAATGTTTTAGAGCGTCGTCAACGTGA


 


TGCGGAAAACAAGAGCCAAGGCAATGTTTTAGAGCGTCGTCAACGTGATGCAGAAAACAGA


 


AGCCAAGGCAATGTTTTAGAGCGTCGTCAACGTGATGCAGAAAACAGAAGCCAAGGCAATG


 


TTCTAGAGCGTCGTCAACGTGATGCAGAAAACAGAAGCCAAGGCAATGTTCTAGAGCGTCG


 


TCAACGTGATGCAGAAAACAGAAGCCAAGGCAATGTTCTAGAGCGTCGTCAACGCGATGCA


 


GAAAACAGAAGCCAAGGTAATGTTCTAGAGCGTCGTCAACGTGATGCAGAAAACAGAAGCC


 


AAGGCAATGTTTTAGAGCGTCGTCAACGTGATGCAGAAAACAGAAGCCAAGGCAATGTTTT


 


AGAGCGTCGTCAACGTGATGCAGAAAACAGAAGCCAAGGCAATGTTTTAGAGCGTCGTCAA


 


CGTGATGCGGAAAACAAGAGCCAAGTAGGTCAACTTATAGGG


 


SEQ ID NO: 9



Construct 3: gbs1309p



AGTGTAACCTATTCACAGTCTGAACGTACGGTTGTTTTCTCTTTTGGAGAAATAACATTTA


 


GTAGGAGTCGCTGGACAAATGGCTTTGAAACTAGAATACCAGTAGATGAGTGGTTAGGTCT


 


TGAAAAATATAAGAGATATTCAATAGAATTCTTATATCATGTTGCAAAATTGGCTACAATG


 


ATGCCTTATCGTCAAGTTTGCAAAGTAATAGATAGCACTTTGCAAACAATCATAACAAAAG


 


ACTGTGTTTTAAAAGCAGTAAAATTTGTAGAAAAATTGTTAAAAGAAAAAGAACGCTATCG


 


TTTTTATTTGGAAGAGCCACCCGAACGTAAAAAAGTGAAAAAACTGTATGTTGAGGGTGAT


 


GGAGTCATGATTAAAAGCACAGATTCTAGAGAGGAAAGAAGGTATTTAGATTTAACACATT


 


TTGTTATTCATACAGGCTCAAAAAAAGTTTCTACTAAAAGATATGAATTGCAGGACAAGCA


 


CGAAATATTACAGCTTAATTATGATAAAGCTAAATATAATCTTTTAGATTATATTTATAAT


 


AACTATGAAGTAGATGACGATACTATTTTAATCACTAACTCTGATATGGGTAAAGGCTATA


 


CTAGTAGAGTTTTTAAGGAATTAGGAAAAGCACTTAAGGTAAAGAAACATGAGCATTTTTG


 


GGATATCTATCATGTTAAAGAAAAGTTAAGTTCATACCTTAGAAAATATCCAATTGAATTA


 


ACCGATTTTGCTTTAGATGCGGTAAAAAAATATAATTCTGATAAGCTTGAATTAGTTTTTG


 


ATACTGTTGAATCACTGATTTGTGATGAACTTGAAGATCAAGAATTTCAGAAGTTTAAGAA


 


AAAAGTATTAAATAATTTCAAATATATAAAACCAGCTCATCTTAGAAATCTTTCAAATCGT


 


GGTATTGGTATCATGGAATCACAACACAGAAAGATAACGTATAGAATGAAGCGACGTGGCA


 


TGTATTGGTCAAAGTGGGGAATCTCCACAATGGCAAATATGATTATACTTGAAAGAGCTAA


 


CGGTTTACGAGAATTATTTTTCGGTTCTTGGAGAAAGGTATACAGTGAGTATAAAGAAGGT


 


TCATTTAGTGCAGGGCGACTTTTTAAAAAGACAGATGAATTAGATAAATTTTCTAAGCCCC


 


TTCTAAAAAATGGCAGAAAATGGAGTATAACAGGAATCAAA


 


SEQ ID NO: 10



Construct 4: gbs1477p



GACGACGTAACAACTGATACTGTGACCTTGCACAAGATTGTCATGCCACAAGCTGCATTTG


 


ATAACTTTACTGAAGGTACAAAAGGTAAGAATGATAGCGATTATGTTGGTAAACAAATTAA


 


TGACCTTAAATCTTATTTTGGCTCAACCGATGCTAAAGAAATTAAGGGTGCTTTCTTTGTT


 


TTCAAAAATGAAACTGGTACAAAATTCATTACTGAAAATGGTAAGGAAGTCGATACTTTGG


 


AAGCTAAAGATGCTGAAGGTGGTGCTGTTCTTTCAGGGTTAACAAAAGACACTGGTTTTGC


 


TTTTAACACTGCTAAGTTAAAAGGAACTTACCAAATCGTTGAATTGAAAGAAAAATCAAAC


 


TACGATAACAACGGTTCTATCTTGGCTGATTCAAAAGCAGTTCCAGTTAAAATCACTCTGC


 


CATTGGTAAACAACCAAGGTGTTGTTAAAGATGCTCACATTTATCCAAAGAATACTGAAAC


 


AAAACCACAAGTAGATAAGAACTTTGCAGATAAAGATCTTGATTATACTGACAACCGAAAA


 


GACAAAGGTGTTGTCTCAGCGACAGTTGGTGACAAAAAAGAATACATAGTTGGAACAAAAA


 


TTCTTAAAGGCTCAGACTATAAGAAACTGGTTTGGACTGATAGCATGACTAAAGGTTTGAC


 


GTTCAACAACAACGTTAAAGTAACATTGGATGGTAAAGATTTTCCTGTTTTAAACTACAAA


 


CTCGTAACAGATGACCAAGGTTTCCGTCTTGCCTTGAATGCAACAGGTCTTGCAGCAGTAG


 


CAGCTGCTGCAAAAGACAAAGATGTTGAAATCAAGATCACTTACTCAGCTACGGTGAACGG


 


CTCCACTACTGTTGAAGTTCCAGAAACCAATGATGTTAAATTGGACTATGGTAATAACCCA


 


ACGGAAGAAAGTGAACCACAAGAAGGTACTCCAGCTAACCAAGAAATTAAAGTCATTAAAG


 


ACTGGGCAGTAGATGGTACAATTACTGATGTTAATGTTGCAGTTAAAGCTATCTTTACCTT


 


GCAAGAAAAACAAACGGATGGTACATGGGTGAACGTTGCTTCACACGAAGCAACAAAACCA


 


TCACGCTTTGAACATACTTTCACAGGTTTGGATAATACTAAAACTTACCGCGTTGTCGAAC


 


GTGTTAGCGGCTACACTCCAGAATATGTATCATTTAAAAATGGTGTTGTGACTATCAAGAA


 


CAACAAAAACTCAAATGATCCAACTCCAATCAACCCATCAGAACCAAAAGTGGTGACTTAT


 


GGACGTAAATTTGTGAAAACAAATCAAGCTAACACTGAACGCTTGGCAGGAGCTACCTTCC


 


TTGTTAAGAAAGAAGGAAAATACTTGGCACGTAAAGCAGGTGCAGCAACTGCTGAAGCAAA


 


GGCAGCTGTAAAAACTGCTAAACTAGCATTGGATGAAGCTGTTAAAGCTTATAACGACTTG


 


ACTAAAGAAAAACAAGAAGGCCAAGAAGGTAAAACAGCATTGGCTACTGTTGATCAAAAAC


 


AAAAAGCTTACAATGACGCTTTTGTTAAAGCTAACTACTCATATGAATGGGTTGCAGATAA


 


AAAGGCTGATAATGTTGTTAAATTGATCTCTAACGCCGGTGGTCAATTTGAAATTACTGGT


 


TTGGATAAAGGCACTTATAGCTTGGAAGAAACTCAAGCACCAGCAGGTTATGCGACATTGT


 


CAGGTGATGTAAACTTTGAAGTAACTGCCACATCATATAGCAAAGGGGCTACAACTGACAT


 


CGCATATGATAAAGGATCTGTAAAAAAAGATGCCCAACAAGTTCAAAACAAAAAAGTAACC


 


ATCCCACAAACAGGTGGTATTGGTACA


 


SEQ ID NO: 11



Construct 5: gbs1478p



GAAAGTACCGTACCGGAAAATGGTGCTAAAGGAAAGTTAGTTGTTAAAAAGACAGATGACC


 


AGAACAAACCACTTTCAAAAGCTACCTTTGTTTTAAAAACTACTGCTCATCCAGAAAGTAA


 


AATAGAAAAAGTAACTGCTGAGCTAACAGGTGAAGCTACTTTTGATAATCTCATACCTGGA


 


GATTATACTTTATCAGAAGAAACAGCGCCCGAAGGTTATAAAAAGACTAACCAGACTTGGC


 


AAGTTAAGGTTGAGAGTAATGGAAAAACTACGATACAAAATAGTGGTGATAAAAATTCCAC


 


AATTGGACAAAATCACGAAGAACTAGATAAGCAGTATCCCCCCACAGGAATTTATGAAGAT


 


ACAAAGGAATCTTATAAACTTGAGCATGTTAAAGGTTCAGTTCCAAATGGAAAGTCAGAGG


 


CAAAAGCAGTTAACCCATATTCAAGTGAAGGTGAGCATATAAGAGAAATTCCAGAGGGAAC


 


ATTATCTAAACGTATTTCAGAAGTAGGTGATTTAGCTCATAATAAATATAAAATTGAGTTA


 


ACTGTCAGTGGAAAAACCATAGTAAAACCAGTGGACAAACAAAAGCCGTTAGATGTTGTCT


 


TCGTACTCGATAATTCTAACTCAATGAATAACGATGGCCCAAATTTTCAAAGGCATAATAA


 


AGCCAAGAAAGCTGCCGAAGCTCTTGGGACCGCAGTAAAAGATATTTTAGGAGCAAACAGT


 


GATAATAGGGTTGCATTAGTTACCTATGGTTCAGATATTTTTGATGGTAGGAGTGTAGATG


 


TCGTAAAAGGATTTAAAGAAGATGATAAATATTATGGCCTTCAAACTAAGTTCACAATTCA


 


GACAGAGAATTATAGTCATAAACAATTAACAAATAATGCTGAAGAGATTATAAAAAGGATT


 


CCTACAGAAGCTCCTAGAGCTAAATGGGGATCAACTACAAACGGACTTACTCCAGAGCAAC


 


AAAAGCAGTACTATCTTAGTAAAGTAGGGGAAACATTTACTATGAAAGCCTTCATGGAGGC


 


AGATGATATTTTGAGTCAAGTAGATCGAAATAGTCAAAAAATTATTGTTCATATAACTGAT


 


GGTGTTCCAACAAGATCATATGCTATTAATAATTTTAAATTGGGTGCATCATATGAAAGCC


 


AATTTGAACAAATGAAAAAAAATGGATATCTAAATAAAAGTAATTTTCTACTTACTGATAA


 


GCCCGAGGATATAAAAGGAAATGGGGAGAGTTACTTTTTGTTTCCCTTAGATAGTTATCAA


 


ACACAGATAATCTCTGGAAACTTACAAAAACTTCATTATTTAGATTTAAATCTTAATTACC


 


CTAAAGGTACAATTTATCGAAATGGACCAGTAAGAGAACATGGAACACCAACCAAACTTTA


 


TATAAATAGTTTAAAACAGAAAAATTATGACATCTTTAATTTTGGTATAGATATATCTGCT


 


TTTAGACAAGTTTATAATGAGGATTATAAGAAAAATCAAGATGGTACTTTTCAAAAATTGA


 


AAGAGGAAGCTTTTGAACTTTCAGATGGGGAAATAACAGAACTAATGAAGTCATTCTCTTC


 


TAAACCTGAGTATTATACCCCGATAGTAACTTCATCCGATGCATCTAACAATGAAATTTTA


 


TCTAAAATTCAGCAACAATTTGAAAAGGTTTTAACAAAAGAAAACTCAATTGTTAATGGAA


 


CTATAGAAGATCCTATGGGTGACAAAATCAATTTACAGCTTGGCAACGGACAAACATTGCA


 


ACCAAGTGATTATACTTTACAGGGAAATGATGGAAGTATAATGAAAGATAGCATTGCAACT


 


GGTGGGCCTAATAATGATGGTGGAATACTTAAAGGGGTTAAATTAGAATACATCAAAAATA


 


AACTCTACGTTAGAGGTTTGAACTTAGGGGAGGGACAAAAAGTAACACTCACATATGATGT


 


GAAACTAGATGACAGTTTTATAAGTAACAAATTCTATGACACTAATGGTAGAACAACATTG


 


AATCCTAAATCAGAGGATCCTAATACACTTAGAGATTTTCCAATCCCTAAAATTCGTGATG


 


TGAGAGAATATCCTACAATAACGATTAAAAACGAGAAGAAGTTAGGTGAAATTGAATTTAC


 


AAAAGTTGATAAAGATAATAATAAGTTGCTTCTCAAAGGAGCTACGTTTGAACTTCAAGAA


 


TTTAATGAAGATTATAAACTTTATTTACCAATAAAAAATAATAATTCAAAAGTAGTGACGG


 


GAGAAAACGGCAAAATTTCTTACAAAGATTTGAAAGATGGCAAATATCAGTTAATAGAAGC


 


AGTTTCGCCGAAGGATTATCAAAAAATTACTAATAAACCAATTTTAACTTTTGAAGTTGTT


 


AAAGGATCGATACAAAATATAATAGCTGTTAATAAACAGATTTCTGAATATCATGAGGAAG


 


GTGACAAGCATTTAATTACCAACACGCATATTCCACCAAAAGGAATTATTCCGATGACAGG


 


TGGGAAA


 


SEQ ID NO: 12



Construct 6: gbs2018p



GATACTAGTTCAGGAATATCGGCTTCAATTCCTCATAAGAAACAAGTTAATTTAGGGGCGG


 


TTACTCTGAAGAATTTGATTTCTAAATATCGTGGTAATGACAAAGCTATTGCTATACTTTT


 


AAGTAGAGTAAATGATTTTAATAGAGCATCACAGGATACACTTCCACAATTAATTAATAGT


 


ACTGAAGCAGAAATTAGAAATATTTTATATCAAGGACAAATTGGTAAGCAAAATAAACCAA


 


GTGTAACTACACATGCTAAAGTTAGTGATCAAGAACTAGGTAAGCAGTCAAGACGTTCTCA


 


AGATATCATTAAGTCATTAGGTTTCCTTTCATCAGACCAAAAAGATATTTTAGTTAAATCT


 


ATTAGCTCTTCAAAAGATTCGCAACTTATTCTTAAATTTGTAACTCAAGCCACGCAACTGA


 


ATAATGCTGAATCAACAAAAGCTAAGCAAATGGCTCAAAATGACGTGGCCTTAATAAAAAA


 


TATAAGCCCCGAAGTCTTAGAAGAATATAAAGAAAAAATTCAAAGAGCTAGCACTAAGAGT


 


CAAGTTGATGAGTTTGTAGCAGAAGCTAAAAAAGTTGTTAATTCCAATAAAGAAACGTTGG


 


TAAATCAGGCCAATGGTAAAAAGCAAGAAATTGCTAAGTTAGAAAATTTATCTAACGATGA


 


AATGTTGAGATATAATACTGCAATTGATAATGTAGTGAAACAGTATAATGAAGGTAAGCTC


 


AATATTACTGCTGCAATGAATGCTTTAAATAGTATTAAGCAAGCAGCACAGGAAGTTGCCC


 


AGAAAAACTTACAAAAGCAGTATGCTAAAAAAATTGAAAGAATAAGTTCAAAAGGATTAGC


 


GTTATCTAAAAAGGCTAAAGAAATTTATGAAAAGCATAAAAGTATTTTGCCTACACCTGGA


 


TATTATGCAGACTCTGTGGGAACTTATTTGAATAGGTTTAGAGATAAACAAACTTTCGGAA


 


ATAGGAGTGTTTGGACTGGTCAAAGTGGACTTGATGAAGCAAAAAAAATGCTTGATGAAGT


 


CAAAAAGCTTTTAAAAGAACTTCAAGACCTTACCAGAGGTACTAAAGAAGATAAAAAACCA


 


GACGTTAAGCCAGAAGCCAAACCAGAGGCCAAACCAAATATTCAAGTACCTAAACAAGCAC


 


CTACAGAAGCTGCAAAACCAGCTTTGTCACCAGAAGCCTTGACAAGATTGACTACATGGTA


 


TAATCAAGCTAAAGATCTGCTTAAAGATGATCAAGTAAAGGACAAATACGTAGATATACTT


 


GCAGTTCAAAAAGCTGTTGACCAAGCTTATGATCATGTGGAAGAGGGAAAATTTATTACCA


 


CTGATCAAGCAAATCAATTAGCTAACAAGCTACGTGATGCTTTACAAAGTTTAGAATTAAA


 


AGATAAAAAAGTAGCCAAACCAGAAGCCAAACCAGAGGCCAAACCAGAAGCTAAGCCAGAA


 


GCTAAGCCAGAAGCTAAGCCAGAAGCTAAGCCAGAGGCCAAACCAGAAGCTAAGCCAGACG


 


TTAAGCCAGAAGCTAAACCAGACGTTAAACCAGAGGCTAAGCCAGAAGCTAAACCAGAGGC


 


TAAGTCAGAAGCTAAACCAGAGGCTAAGCTAGAAGCTAAACCAGAGGCCAAACCAGCAACC


 


AAAAAATCGGTTAATACTAGCGGAAACTTGGCGGCTAAAAAAGCTATTGAAAACAAAAAGT


 


ATAGTAAAAAATTACCATCAACG






Expression and Purification of Proteins:

E. coli BL21 Star® cells harboring the recombinant plasmid were grown into log phase in the required culture volume. Once an OD600nm of 0.6 was reached the culture was induced with 0.5 mM IPTG for 3 hours at 37° C. The cells were harvested by centrifugation, lysed by a combination of the freeze-thaw method followed by disruption of cells with ‘Bug-buster®’, (Novagen). The lysate was separated by centrifugation into soluble (supernatant) and insoluble (pellet) fractions. Depending on the location of the protein different purification strategies were applied. A) If the His-tagged protein was in the soluble fraction, protein purification was done by binding the supernatant to Ni-Sepharose beads (Ni-SepharoseTM 6 Fast Flow, GE Healthcare). Due to the presence of the hexa Histidine (6×HIS) at the C terminus of the expressed protein, it bound to the Ni-Sepharose while the other contaminating proteins were washed from the column by wash buffer. The protein was eluted by 500 mM Imidazole in 20 mM NaH2PO4, 0.5 mM NaCl buffer at pH 7.4. The eluate was concentrated, assayed by Bradford for protein concentration and checked by SDS-PAGE and Western blot. B) If the protein was present in the insoluble fraction, the pellet was solubilized in suitable buffer containing 8 M urea and applied onto the Ni-NTA column under denaturing conditions (in buffer containing 8 M urea) using the same materials and procedure as mentioned above. Contaminating proteins were washed from the column by wash buffer without urea. Refolding of the His-tagged protein was performed while the protein was immobilized on the Ni-NTA matrix. After renaturation, proteins were eluted by the addition of 500 mM Imidazole. The eluate was dialyzed to remove traces of urea and concentrated if the volume was large, checked by SDS-PAGE and measured by the Bradford method.
Animal Protection Studies
Animals:
CD-1 female mice (6-8 weeks) were used for these studies.
Active Immunization, Generation of Hyper-Immune Mouse Sera:
50 μg of recombinant protein was injected subcutaneously into CD-1 mice, adjuvanted with Complete Freund's adjuvant (CFA). Animals were boosted twice with the same amount of protein and Incomplete Freund's adjuvant (IFA) at days 14 and 28. The published protective Sip (gbs0031) protein antigen (Brodeur et al., Infect Immun 68(10):5610-5618 (2000)) was used as a positive control, while mice immunized with adjuvant only served as negative controls. Antibody titres were measured at day 35 by ELISA using the respective recombinant proteins. In case of hyper-immune sera generation mice were terminally bled at day 35.
Generation of Hyperimmune Rabbit Sera:
Polyclonal rabbit sera were generated for gbs0031, gbs0233p, gbs1087p, gbs1309p, gbs1477p, gbs1478p and gbs2018p at Charles
River Laboratories, Kislegg, Germany. 250 μg of recombinant protein was injected into New Zealand White rabbits, adjuvanted with Complete Freund's adjuvant (CFA) Animals were boosted three times with the same amount of protein, but with Incomplete Freund's adjuvant (IFA) at days 28, 42 and 56. Antibody titers were measured at day 38 and 52 by ELISA using the respective recombinant proteins. Rabbits were terminally bled at day 70.
Passive Immunization:
CD-1 mice were immunized intraperitoneally 1 to 3 hours before the bacterial challenge with 150 μl mouse or rabbit hyperimmune sera.
Bacterial Challenge:
Freshly grown S. agalactiae strains C388/90 (serotype Ia/c), A909 (serotype Ia/c), ATCC12401 (serotype Ib), ATCC12403 (serotype III), COH1 (serotype III), BAA22 (serotype III), 2603V/R (serotype V), ATCC49447 (serotype V), BAA23 (serotype V) were used for animal challenge studies. In order to determine the viable cell numbers present in the bacterial inoculum, cfus were determined via plating on blood agar plates. 106-108 cfus were applied intraperitoneally into mice. Protection by immunization was measured by a lethal sepsis model, where survival rates were followed for 1 to 2 weeks post-challenge and survival was expressed as percentage of the total number of animals (10 mice/group).
Results
By using a genomic scale antigen identification method we selected Group B streptococcal antigens based on immunogenicity in humans (WO2004/099242) and pre-selected vaccine candidates based on in vitro assays. Here we show immune protection by six Group B streptococcal antigens in animal models. The first screening model was set up using adult mice and the mouse-adapted S. agalactiae ATCC12403 serotype III strain that was also used for the genomic library construction and cloning of some of the vaccine candidates. We set up the method with CD-1 mice and defined the LD90-LD100 dose. The model set up was further optimized by using positive and negative control sera. Protection was estimated by reduced lethality of mice immunized with Sip or anti-Sip immune sera relative to animals immunized with adjuvant alone or treated with control sera. Based on these data, CD-1 mice and a challenge dose between 5×107 to 1×108 cfu was used for further studies. Mice were immunized first with the recombinant antigens adjuvanted with CFA/IFA and in subsequent experiments with hyper-immune mouse sera transferred to naüve animals before challenge with S. agalactiae ATCC12403 (serotype III). In the active, as well as in the passive model, several protective antigens were identified that showed variable protection levels, ranging from higher, equal or lower survival relative to Sip. Since several different Group B Streptococcus serotypes are able to cause severe disease in humans, it is important to test cross-protection of vaccine candidates against all major serotypes in animal experiments. Moreover, it has been firmly demonstrated that protective antigens show strain-dependent variations not only in their primary sequences and expression, but also in their protective capacity. For that reason, we have set up the screening model with several different S. agalactiae strains representing the major serotypes, Ia, Ib, III and V. Strain-dependent protection within one serotype was also addressed by using 2-3 different strains of the most common serotypes Ia, III and V. In order to perform this large number of experiments with the minimal animal sacrifice and good comparability, we generated hyper-immune rabbit sera for all in vitro selected recombinant antigens. Three rabbits were immunized with each individual antigen adjuvanted with CFA/IFA using a standard protocol. Animals were pre-screened for pre-existing GBS-specific antibodies by testing their sera with ELISA and only animals without a significant reaction were included in immunization studies. The individual hyper-immune sera were then analyzed for antigen-specific antibody levels and used in pools for further analyses. Thus, the very same immune sera were used for passive protection studies with nine different GBS strains that we found useful for animal studies. As a result of these experiments we could identify six novel vaccine candidates—gbs0233p, gbs1087p, gbs1309p, gbs1477p, gbs1478p and gbs2018p—that showed protection against at least one serotype when used as a sole antigen (FIG. 1).
In order to examine benefits of combinations of different antigenic components, we performed passive protection studies by combining rabbit sera with different antigen specificities. With different combinations using these six protective vaccine candidates, we could demonstrate increased protection compared to the single proteins against all the tested GBS serotypes. The combination of gbs1477p+gbs2018p provided a significantly increased level of protection against many serotypes. The best protection seen so far was achieved with a combination of gbs1087p+gbs1477p+gbs1478p+gbs2018p that protected most of the mice against all nine tested GBS strains (FIG. 2).
Example 2
Surface Exposure and Induction of Functional Antibodies by Group B Streptococcal Antigens
Experimental Procedures
FACS Analysis:
The S. agalactiae strain to be tested was inoculated from a glycerol stock into 5 ml THB medium and incubated over night at 37° C. The overnight culture was reinoculated by adding 200 μl into 10 ml fresh THB medium and incubated until an OD600nm of approximately 1 was reached ( ̃5×108 cells/ml). The bacterial cells were pelleted by centrifugation at 4,000 rpm for 5 min and washed twice with 2 ml HBSS. The final pellet was resuspended in HBSS with 1% BSA to give a cell density of 5×106 cells/ml. To 100 μl bacterial suspension 1 μl serum was added and incubated for 45 min on ice. Bacteria were pelleted by centrifugation at 1,000 g for 4 min and washed once with 150 μl HBSS with 1% BSA and resuspended in 100 μl HBSS with 1% BSA. To the opsonised bacteria 1 μl of the secondary antibody (goat F(ab)2 fragment anti rabbit IgG coupled with PE) was added and incubated for 45 min on ice in the dark. The cells were washed twice with 150 μl HBSS as described above and dissolved in 250 μl HBSS, the cells were fixed by the addition of 250 μl 4% para-formaldehyde. The fluorescent staining of the bacteria was measured by FACS analysis.
Opsonophagocytic Killing Assay
Preparation of Bacterial Cells:
The S. agalactiae strain to be tested was inoculated from a glycerol stock into 5 ml THB medium and incubated overnight at 37° C. The over night culture was reinoculated by adding 200 μl into 10 ml fresh THB medium and incubated until an OD600nm of approximately 1 was reached. The bacteria were pelleted by centrifugation at 4,000 rpm for 5 min and washed twice with 2 ml HBSS. The final pellet was resuspended in HBSS with 0.125% BSA to give a final concentration of 5×104 cells/85 μl.
Preparation of RAW264.7 Cells:
Cells were cultivated in T175 flasks with 25 ml DMEM high glucose medium at 37° C. with 5% CO2. Cells were detached from the flasks by scraping and collected by low speed centrifugation at 1,000 rpm for 10 min and washed twice with 50 ml HBSS with 10 mM glucose and resuspended in HBSS with 10 mM glucose to give a cell concentration of 1×107 cells/ml.
Opsonophagocytic Killing Assay:
Bacteria (85 μl) were mixed with 10 μl guinea pig complement and 5 μl prediluted serum and incubated for 60 min at 6° C. with shaking (500 rpm). To the opsonised bacteria 100 μl (1×106 cells) RAW264.7 cells were added. Three aliquots of 10 μl were taken and each added to 1.5 ml water after 5 min incubation, 100 μl were plated on blood agar plates to determine the initial bacterial count, T0. The suspensions with opsonised bacteria and RAW264.7 cells were incubated for one hour at 37° C. with shaking (500 rpm). After 60 min incubation three aliquots of 10 μl were removed and each diluted in 1.5 ml, after 5 min incubation, 100 μl were plated on blood agar plates to determine T60. After overnight cultivation cfus were determined with a colony counter.
Evaluation:
For each sample the relationship between the cfu at T0 and T60 was determined. The percentage killing of each test serum was related to the respective preimmune serum using the relationship between T0 and T60 with the formula 100-100×(test serum/preimmune serum).
Results
The analyses of surface expression of gbs0233, gbs1087, gbs1309, gbs1477, gbs1478 and gbs2018 have been performed by FACS analysis using the very same pooled rabbit hyperimmune sera that were tested for protection in animal studies. These six protective antigens were detected on the surface of Group B streptococcal strains. Four of the antigens (gbs1087, gbs1477, gbs1478, gbs2018) were most consistently detected (FIG. 3), gbs0233 was not expressed in vitro by all strains and gbs1309 was mainly detected in the bacterial supernatant. The in vitro expression experiments have been performed with nine different strains from the serotypes Ia, Ib, II, III, IV and V; the most comprehensive studies have been performed with the serotype III strain ATCC12403.
Based on the passive protection data, it is firmly established that protection by the selected six vaccine candidates is mainly mediated by antibodies. The ability to measure functional antibodies in in vitro assays is essential for the development of both a prophylactic vaccine and an antibody-based therapy or prevention. Nine different S. agalactiae strains representing six serotypes (Ia, Ib, II, III, IV and V) were used to evaluate gbs0233p, gbs1087p, gbs1309p, gbs1477p, gbs1478p and gbs2018p for induction of functional antibodies. Included in the opsonophagocytic killing assays were two GBS strains representing serotypes II and IV that were not suitable for animal testing. As an example of the in vitro assays, results with two strains are presented in FIG. 4, the serotype II strain AC3912 (FIG. 4A) and the serotype V strain BAA23 (FIG. 4B). Simultaneously with the opsonophagocytic killing assay cells were tested for in vitro expression of the tested antigens by Western blot and FACS analysis. At a serum dilution of 1:200, only gbs1478p showed more than 50% killing of the serotype II strain AC3912 (FIG. 4A), both gbs1477p and gbs1478p showed more than 50% killing of the serotype V strain BAA23 (FIG. 4B). The remaining antigens showed less than 50% killing of the strains tested in FIG. 4, which in most cases can be explained by poor in vitro expression of the antigens in these strains.
Example 3
Sequence Conservation of Protective Group B Streptococcal Antigens
Experimental Procedures
Sequence Analyses of S. Agalactiae Genes:
In order to determine the sequence of an antigen from diverse S. agalactiae strains, PCR was performed with primers specific for the gene of interest. S. agalactiae strains used for these analyses are shown in Tables 2 and 13. Oligonucleotide sequences as primers for PCR were designed for the selected antigens in order to be able to amplify the full gene. Sequencing was performed with dedicated primers using the PCR products as templates. The sequences of the oligonucleotides are listed in Table 3. Genomic DNA of all S. agalactiae strains was prepared as described in WO2004/099242. PCR was performed in a reaction volume of 25 μl using Taq polymerase (1 U), 200 nM dNTPs, 10 pMol of each oligonucleotide and the kit according to the manufacturer's instructions (Invitrogen, The Netherlands). As standard, 30 cycles (1×: 5 min 95° C., 30×: 30 sec. 95° C., 30 sec. 56° C., 30 sec. 72° C., 1×: 4 min 72° C.) were performed, unless conditions had to be adapted for individual primer pairs. PCR samples were sequenced with the oligonucleotides as listed in Table 3.
Results
The genomic sequence of eight individual strains of S. agalactiae (Tables 2 and 4) has been published and was compared for the six antigens shown to be protective under Example 1. The comparison showed that the proteins gbs0233 and gbs1087 are highly conserved (more than 99 and 91% identity, respectively; Tables 2 and 4), although gbs1087 displayed various numbers of repeats in the different GBS strains (see also WO2004/035618). This high degree of protein sequence identity (gbs0233: >99%; gbs1087: >86%) could also be observed for the strains that were subjected to DNA sequence analyses as listed in Tables 5, 7, 8, and Table 13 and in the Sequence listing. The gbs0233 protein from any of the analyzed strains showed at least 98.7% amino acid sequence identity to gbs0233 from S. agalactiae NEM316, with only 6 amino acid position showing a change. The sequences of the gbs1087 proteins from the analyzed strains were also highly conserved, yet the different strains harboured between a single and up to 29 repeats of a highly conserved 17 amino acid long sequence. The sequences of proteins gbs1309 and gbs2018 showed high sequence conservation in 7 genomic strains (more than 87 and 77% identity, respectively), while protein sequences diverged more significantly in strain COH1 (69.9 and 47.7%, respectively; Table 4). The gbs1309 protein showed a similar high degree of amino acid sequence identity (89.6%) in the sequenced GBS strains (Table 5, 9, 13 and Sequence listing), while the gbs2018 protein can be classified in two clades, with 95% of strains belonging to one clade with at least 60.8% sequence identity and 3 strains COH1(III), BAA22(III) and 49447(V) belonging to the second clade. The protein gbs 1478 is highly conserved in 6 genomic strains (more than 87% identity), yet the strains COH1 and A909 show a lower amino acid sequence identity of approximately 43% (Table 4). Protein gbs1478 is conserved in most analyzed GBS strains as shown in Table 5, 11, 13 and the Sequence listing, but exists as 2 distinct clades with an amino acid sequence identity of more than 80% in the dominant clade (approx. 80% of analyzed strains) and more than 99% in the second clade. The protein gbs1477 shows the highest degree of amino acid sequence variability, with six distinct clades that can be characterized. Strains COH1 and A909 do not encode a homologous protein with significant amino acid sequence identity (Table 4). The sequence analyses of the gbs1477 gene from further distinct GBS strains revealed that all selected strains encode a protein homologous to gbs1477 and that all six clades were covered by these sequences (Table 5, 6, 10, 13, Sequence listing and FIG. 5). The prototype sequences for the 6 clades of gbs 1477 are: strain 12401 (clade 1; SEQ ID NO: 93), strain IC254 (clade 2; SEQ ID NO: 110), strain 126H4A (clade 3; SEQ ID NO: 94), strain 49447 (clade 4; SEQ ID NO: 95), strain C388/90 (clade 5; SEQ ID NO: 100) and strain NEM316 (clade 6; SEQ ID NO: 223 and SEQ ID NO: 361); (for all sequences, see Sequence listing). Within any single clade the level of amino acid sequence identity reaches at least 98%.
Example 4
Group B Streptococcal Antigens and Combinations Thereof as Well as Mouse Monoclonal Antibodies, Generated Against these Antigens, Induce Protective Immune Responses Against Lethal Sepsis in an i.p. Challenge Model
Experimental Procedures
Cloning and Expression of Recombinant Group B Streptococcal Proteins
Cloning of Genes:
The gene of interest was amplified from genomic DNA of S. agalactiae ATCC12403 (serotype III) by PCR using gene specific primers. Apart from the gene specific part, the primers had restriction sites that aided in a directional cloning of the amplified PCR product. The gene annealing (specific) part of the primer ranged between 15-30 bases in length. The PCR products obtained were digested with the appropriate restriction enzymes and cloned into the pET28b (+) vector (Novagen) for His-tagged proteins. Once the recombinant plasmid was confirmed to contain the gene of interest, E. coli BL21 Star® cells (Invitrogen) that served as expression host were transformed. Cloning of the gbs1087, gbs1477 and gbs1478 genes has been performed using genomic DNA from strain S. agalactiae 6313 (serotype III) in the vector pET28a (+). The constructs of the selected antigens are listed in Table 1.
Expression and Purification of Proteins:
E. coli BL21 Star® cells harboring the recombinant plasmid were grown into log phase in the required culture volume. Once an OD600m of 0.6 was reached the culture was induced with 0.5 mM IPTG for 3 hours at 37° C. The cells were harvested by centrifugation, lysed by a combination of the freeze-thaw method followed by disruption of cells with ‘Bug-buster®’ (Novagen). The lysate was separated by centrifugation into soluble (supernatant) and insoluble (pellet) fractions. Depending on the location of the protein different purification strategies were applied. A) If the His-tagged protein was in the soluble fraction, protein purification was done by binding the supernatant to Ni-Sepharose beads (Ni-SepharoseTM 6 Fast Flow, GE Healthcare). Due to the presence of the hexa Histidine (6×HIS) at the C terminus of the expressed protein, it bound to the Ni-Sepharose while the other contaminating proteins were washed from the column by wash buffer. The protein was eluted by 500 mM Imidazole in 20 mM NaH2PO4, 0.5 mM NaCl buffer at pH 7.4. The eluate was concentrated, assayed by Bradford for protein concentration and checked by SDS-PAGE and Western blot. B) If the protein was present in the insoluble fraction, the pellet was solubilized in suitable buffer containing 8 M Urea and applied onto the Ni-NTA column under denaturing conditions (in buffer containing 8 M Urea) using the same materials and procedure as mentioned above. Contaminating proteins were washed from the column by wash buffer without urea. Refolding of the His-tagged protein was performed while the protein was immobilized on the Ni-NTA matrix. After renaturation, proteins were eluted by the addition of 500 mM Imidazole. The eluate was dialyzed to remove traces of urea and concentrated if the volume was large, checked by SDS-PAGE and measured by the Bradford method.
Animal Protection Studies
Animals:
CD-1 female mice (6-8 weeks) were used for these studies.
Active Immunization:
25 μg of recombinant protein was injected subcutaneously into CD-1 mice, adjuvanted with ALUM 1% Animals were boosted twice with the same amount of protein and ALUM 1% at days 14 and 28. The published protective Sip (gbs0031) protein antigen was used as a positive control, while mice immunized with adjuvant only served as negative controls. Antibody titres were measured at day 35 by ELISA using the respective recombinant proteins.
Generation of Hyperimmune Rabbit Sera:
Polyclonal rabbit sera were generated for gbs0031, gbs0233p, gbs1087p, gbs1309p, gbs1477p, gbs1478p and gbs2018p at Charles River Laboratories, Kislegg, Germany. 250 μg of recombinant protein was injected into New Zealand White rabbits, adjuvanted with Complete Freund adjuvant (CFA). Animals were boosted three times with the same amount of protein, but with Incomplete Freund adjuvant (IFA) at days 28, 42 and 56. Antibody titers were measured at day 38 and 52 by ELISA using the respective recombinant proteins. Rabbits were terminally bled at day 70.
Generation of Mouse Monoclonal Antibodies:
Monoclonal mouse antibodies were generated against gbs0233p, gbs1087p, gbs1477p, gbs1478p and gbs2018p at Abgent, San Diego, USA. 100 μg of recombinant protein was injected into Balb/c mice, adjuvanted with Complete Freund adjuvant (CFA) Animals were boosted with 50 μg protein and CFA at week 2; at week 3 animals were boosted with the same amount of protein, but with Incomplete Freund adjuvant (IFA) and at week 4 and 5 animals were boosted with 50 μg protein in PBS (without adjuvant). Antibody titers were measured in week 5 by ELISA and Western blotting using the respective recombinant proteins. Spleen cells from mouse with the best titer were fused with myeloma cell F0 using PEG protocol. Subsequently growing fused hybridoma clones were screened against the respective antigen for test of their specificity and sensitivity. ELISA positive clones were tested also by Western blot. Selected clones from this test were subcloned at least two times and antibodies were purified by protein G affinity chromatography from culture medium.
Passive Immunization (Neonates):
Pregnant CD-1 mice were given 0.5 ml undiluted rabbit hyper-immune sera by intraperitoneal injection 2 to 4 days before delivery. Within 48 h after birth, pups were challenged intraperitoneally.
Bacterial Challenge:
Freshly grown S. agalactiae strains C388/90 (serotype Ia/c), A909 (serotype Ia/c), ATCC12401 (serotype Ib), ATCC12403 (serotype III), COH1 (serotype III), ATCCBAA22 (serotype III), 2603V/R (serotype V), ATCC49447 (serotype V), ATCCBAA23 (serotype V) were used for animal challenge studies. In order to determine the viable cell numbers present in the bacterial inoculum, cfus were determined via plating on blood agar plates. 106-108 cfus were applied intraperitoneally into mice. Protection by immunization was measured by a lethal sepsis model, where survival rates were followed for 1 to 2 weeks post-challenge and survival was expressed as percentage of the total number of animals (10 mice/group for active immunization; for neonatal challenge number of animals depends on the litter size).
Results
By using a genomic scale antigen identification method we selected Group B antigens based on immunogenicity in humans (WO04/099242) and pre-selected vaccine candidates based on in vitro assays. We have shown previously immune protection by six Group B streptococcal antigens in animal models. Additionally, with different combinations using these six protective vaccine candidates, we demonstrated increased protection compared to the single proteins against all the tested GBS serotypes. The combination of gbs1477p+gbs2018p provided a significantly increased level of protection against many serotypes. The best protection seen so far was achieved with a combination of gbs1087p+gbs1477p+gbs1478p+gbs2018p that protected most of the mice against all nine tested GBS strains (see Example 1 and FIG. 2). So far these experiments were obtained in serum transfer experiments. We now further substantiated these results by active immunization of mice with two, three or four recombinant proteins using ALUM as adjuvant (FIG. 6). Immunization with single proteins verified the data already obtained with the hyper-immune rabbit sera (FIG. 7).
Since GBS sepsis affects mainly newborns, we have also developed a model that can demonstrate protection in neonatal mice. We established a murine model with passive immunization of pregnant mice with hyperimmune rabbit sera (500 μl i.p.) 2-4 days before delivery and challenging their babies with S. agalactiae 24-48 h after birth. We observed excellent protection of newborn mice born to mothers immunized with the combination of gbs1087p+gbs1477p+gbs1478p+gbs2018p specific immune sera (4×125 μl) (FIG. 8). Hyperimmune sera against the individuals antigens were also effective in this, but overall was lower than that obtained with the combination of four (data not shown). These findings are very significant, since the models with the different GBS strains were very to stringent, resulting in death of infected pups within 24 hours.
Since the protein gbs 1477 has the highest sequence variability and exists in different clades (Table 5 & 6, FIG. 5) the protection was analyzed using the adult sepsis/lethality model. Mice were immunized with fragments (corresponding to gbs1477p of strain 6313) of six different gbs1477 proteins, originated from distinct clades. Protection was measured against the homologous as well as against the heterologous clade (FIG. 9). The best protection was always obtained when immunization and challenge is done with the homologous clade. The more variable the sequences of the different clades are the lower the protection obtained in the sepsis model.
This invention includes also protection data by mouse monoclonal antibodies. mAbs were generated against gbs0233p, gbs1087p, gbs1477p, gbs1478p and gbs2018p. Selection of hybridoma supernatants were performed using antigen-specific ELISA and/or FACS analysis. Per antigen two mAbs were selected and tested in the passive transfer model using 50 μg purified mAb. We demonstrate in this invention that we obtain protection with a single mAb against at least one serotype (FIG. 10). In order to examine benefits of combinations of different mAb components, we performed passive protection studies by combining mAbs with different antigen specificities. We could demonstrate increased protection compared to the single mAbs against all the tested GBS serotypes. The best protection seen so far was achieved with a combination of mAbs against gbs0233p+gbs1087p+gbs1477p+gbs1478p+gbs2018p that protected most of the mice against all nine tested GBS strains (FIG. 11).
Example 5
Group B Streptococcal Antigens and Mouse Monoclonal Antibodies, Generated Against these Antigens, Induce Functional Antibodies Against Group B streptococcus 
Experimental Procedures
FACS Analysis
The Streptococcus agalactiae strain to be tested was inoculated from a glycerol stock into 5 ml THY medium and incubated over night at 37° C. The overnight culture was re-inoculated by adding 200 μl into 10 ml fresh THY medium and incubated until an OD600nm of approximately 1 was reach ( ̃5×108 cells/ml). The bacteria were pelleted by centrifugation at 4,000 rpm for 5 min and washed twice with 2 ml HBSS. The final pellet was resuspended in HBSS with 1% BSA to give a cell density of 5×106 cells/ml. To 100 μl bacteria, 1 μl immune serum was added and incubated for 45 min on ice. Bacteria were pelleted by centrifugation at 1,000 g for 4 min and washed once with 150 μl HBSS with 1% BSA and resuspended in 100 μl HBSS with 1% BSA. To the opsonised bacteria, 1 μl of the secondary antibody (goat F(ab)2 fragment anti rabbit IgG coupled with PE) was added and incubated for 45 min on ice in dark. The cells were washed twice with 150 μl HBSS as described above and dissolved in 250 μl HBSS, the cells were fixed by addition of 250 μl 4% para-formaldehyde. The fluorescent staining of the bacteria was measured by flow cytometry.
Opsonophagocytic Killing Assay
Preparation of Bacterial Cells:
The Streptococcus agalactiae strain to be tested was inoculated from a glycerol stock into 5 ml THY medium and incubated overnight at 37° C. The over night culture was re-inoculated by adding 200 μl into 10 ml fresh THY medium and incubated until an OD600nm of approximately 1 was reached. The bacteria were pelleted by centrifugation at 4,000 rpm for 5 min and washed twice with 2 ml HBSS. The final pellet was re-suspended in HBSS with 0.125% BSA to give a final concentration of 5×104 cells/85 μl.
Preparation of RAW264.7 Cells:
Cells were cultivated in T175 flasks with 25 ml DMEM high glucose medium at 37° C. with 5% CO2. Cells were detached from the plates by scraping and collected by low speed centrifugation at 1,000 rpm for 10 min and washed twice with 50 ml HBSS with 10 mM glucose and re-suspended in HBSS with 10 mM glucose to give a cell concentration of 1×107 cells/ml.
Opsonophagocytic Killing Assay Conditions:
Bacterial cells (85 μl) were mixed with 10 μl guinea pig complement and 5 μl pre-diluted serum and incubated for 60 min at 6° C. with shaking (500 rpm). To the opsonised bacteria, 100 μl (1×106 cells) of RAW264.7 cells were added. Three aliquots of 10 μl were taken out and added to 1.5 ml water after 5 min incubation, 100 μl were plated out on blood agar plates. This CFU determination served as the initial bacterial count, T0. The suspension of opsonised bacteria and RAW264.7 cells was further incubated at 37° C. with shaking (500 rpm) for 60 min and then the T60 was determined as described for the T0. Blood agar plates were incubated overnight and the CFUs determined on the next day using a colony counter.
Evaluation:
For each serum, the relationship between the CFUs at T0 and T60 was determined for the pre-immune and the immune serum. The percentage of killing of each immune serum was determined by the following formula: 100-100×(immune serum/preimmune serum). A reaction without sera was included in each assay as negative (complement) control.
Results
Based on the passive protection data, it is firmly established that protection by the selected six vaccine candidates is mainly mediated by antibodies. The ability to measure functional antibodies in in vitro assays is essential for the development of both a prophylactic vaccine and an antibody-based therapy or prevention. The same opsonophagocytic killing assay that was developed for the in vitro validation and used for selection of vaccine candidates was employed to analyze the hyperimmune rabbit sera for the presence of functional antibodies. Seven mouse monoclonal antibodies and four rabbit sera representing four antigens were tested in the opsonophagocytic killing assay for functional antibodies and staining in flow cytometry of the serotype III GBS strain ATCC12403 (FIGS. 12 and 13). All monoclonal antibodies bound to the serotype III GBS strain ATCC12403 as measured by flow cytometry (FIG. 12). The opsonophagocytic killing assay with the rabbit and mouse monoclonal antibodies are shown in FIG. 13. At a 1:1,000 dilution of the rabbit sera only gbs1477#28 and gbs1478#4 showed a high killing activity and gbs1087#12 showed low killing activity. The mouse monoclonal antibodies generated against gbs1477p and gbs1478p showed high killing activity and those against gbs1087p and gbs2018p showed only a killing activity at the higher concentration.







TABLE 1







List of genes selected for expression.


The nomenclature of the proteins is derived from the genome of NEM316 (ATCC12403).


The strain and position (start/stop) of the amplicon within the full


length gene/protein are indicated.

















Full


nt
SEQ
aa
SEQ


Construct
Gene
length
Strain
Vector
(start/stop)
ID NO
(start/stop)
ID NO


















1
gbs0233
308
12403
pET28b
37-921
7
13-307
1


2
gbs1087
442
6313
pET28a
106-1062
8
36-354
2


3
gbs1309
403
12403
pET28b
 4-1203
9
 2-401
3


4
gbs1477
674
6313
pET28a
 88-1944
10
30-648
4


5
gbs1478
901
6313
pET28a
 88-2595
11
30-865
5


6
gbs2018
643
12403
pET28b
106-1836
12
36-612
6
















TABLE 2







List of S. agalactiae strains which were used for the first


run of sequencing of the most of the genes of interest, and for


which the genome has been published. (A complete list of strains


used for the first and second run can be found in Table 13.)










S. agalactiae strain
Serotype











GBS strains used for sequencing










IC105
IV



IC458
Ia



0176H4A
II



ATCC12401
Ib



BAA23
V



COH1
III



ATCC12403
III



6313
III







Published genomic GBS strains (Maione et al.,


2005, Science 309(5731): 148-50)










H36B
Ib



COH1
III



CJB111
V



A909
Ia/c



515
Ia



2603V/R
V



NEM316 (ATCC12403)
III



18RS21
II

















TABLE 3







Oligonucleotides used for PCR and sequencing















SEQ


ORF-



ID


protein
PCR
Sequencing Primer
Name
NO:





gbs0233
ICC5455 &
GGGGGCACAATTCCTGTTAT
ICC 5455
13



ICC5456
AAAAAGTGGTGGATAAATTGTTCT
ICC 5456
14


 


gbs1087
ICC5489 &
CATTGTAAATCTTAATGTTAGTATGA
ICC 5489
15



ICC5494
TGACTTTGATTTCCAACACTATCC
ICC 5490
16




GGTTTTAGAACTTGGAAATCAGGA
ICC 5491
17




GGTCTATTAGCTACATTAGTAACCTG
ICC 5492
18




AGAGAAAATAATCACTCTAGTCAAGG
ICC 5493
19




AAAAAGTCACCCTAACCAACC
ICC 5494
20


 


gbs1309
ICC5465 &
AATCATCGGTGAAGTGACGA
ICC 5465
21



ICC5468
CGGTTAATTCAATTGGATATTTTCT
ICC 5466
22




ACTCTGATATGGGTAAAGGCTAT
ICC 5467
23




CTTGAATTATTCTTAAAAAGACCAAAA
ICC 5468
24



ICC5469 &
CCAGTAGATGAGTGGTTAGGTCTTG
ICC 5469
25



ICC5470
AAGATGAGCTGGTTTTATATATTTG
ICC 5470
26


 


gbs1477
ICC5471 &
TTGCAGGTGGAATTTATATTTGG
ICC 5471
27



ICC5479
TTCTTATCTACTTGTGGTTTTGTTTCA
ICC 5477
28




TCTTGGCTGATTCAAAAGCA
ICC 5478
29




GGTTCTGATGGGTTGATTGG
ICC 5479
30



ICC5480 &
AATGGCTCTTGCTTATGATCT
ICC 5472
31



ICC5472
TGTTAGCGGCTACACTCCAG
ICC 5480
32



ICC5471 &
TTGCAGGTGGAATTTATATTTGG
ICC 5471
33



ICC5475
CAACTTTTGGTTCAGTTGG
ICC 5475
34




CCCATTGTCAAACCATTT
ICC 5473
35




GCTACTGCTGAAATCGGTMA
ICC 5474
36



ICC5476 &
CATACATGATCTCAGAACGT
ICC 5476
37



ICC5472
AATGGCTCTTGCTTATGATCT
ICC 5472
38


 


gbs1478
ICC5481 &
TCTAGGATATTCTGTATCTGATCTTAG
ICC 5481
39



ICC5484
CCATCAAAAATATCTGAACCA
ICC 5482
40




GAGGGAACATTATCTAAACGTATTTCA
ICC 5483
41




TTCAATTTTTGAAAAGTACCATCTTG
ICC 5484
42



ICC5485 &
GAACATGGAACACCAACCAA
ICC 5485
43



ICC5488
TCAATTTCACCTAACTTCTTCTCG
ICC 5486
44




TTTTCCAATCCCTAAAATTCG
ICC 5487
45




TTTTCATTTCTATCTCCTTCTTATTC
ICC 5488
46


 


gbs2018
ICC5457 &
AAAAGGCAAAGTTCTGATGAGG
ICC 5457
47



ICC5460
AAAAATGCTTGATGAAGTCAAAA
ICC 5458
48




GTTTGGCTTCTGGCTTAACG
ICC 5459
49




TGATCAAGAACTAGGTAAGCAGTCA
ICC 5460
50



ICC5461 &
CAAATTTAAGAATAAGTTGCGAATC
ICC 5461
51



ICC5462
AGAGTAAATGATTTTAATAGAGCATCA
ICC 5462
52



ICC5463 &
AAAATATTTCTAATTTCTGCTTCAGT
ICC 5463
53



ICC5464
AATTAAAATAAACGTGGTCCTATCC
ICC 5464
54
















TABLE 4





Sequence identity of proteins in published genomes of 8 GBS strains.


ORF indicates the name of the respective gene/protein in the genomic


GBS strain. % Id, amino acid sequence identity in percentage. All comparisons


are performed to the respective protein of GBS strain NEM316.




















S. agalactiae
S. agalactiae
S. agalactiae
S. agalactiae


ORF name
H36B
COH1
CJB111
A909















(NEM316)
ORF
% Id
ORF
% Id
ORF
% Id
ORF
% Id





gbs0233
SAI_0243
99.0
SAN_0021
99.7
SAM_0244
99.0
SAK_0301
99.0


gbs1087
SAI_2325
93.9
SAN_1174
93.3
SAM_1069
91.5
SAK_1142
92.8


gbs1309
SAI_1330
99.4
SAN_1370
69.9
SAM_1259
100.0
SAK_1321
99.7


gbs1477
SAI_1511
47.0
none

SAM_1372
98.4
none


gbs1478
SAI_1512
87.7
SAN_0702
43.2
SAM_1373
99.1
SAK_0780
43.3


gbs2018
SAI_2103
78.8
SAN_2207
47.7
SAM_1974
77.6
SAK_1999
92.3
















S. agalactiae
S. agalactiae
S. agalactiae



ORF name
515
2603V/R
18RS21















(NEM316)
ORF
% Id
ORF
% Id
ORF
% Id







gbs0233
SAL_0280
99.7
SAG0242
99.0
SAJ_0320
99.0



gbs1087
SAL_1159
92.0
SAG1052
100.0
SAJ_1090
100.0



gbs1309
SAL_1364
87.7
SAG1237
99.7
SAJ_2108
90.1



gbs1477
SAL_1486
75.9
SAG1407
47.3
SAJ_1416
47.3



gbs1478
SAL_1487
97.0
SAG1408
97.3
SAJ_1417
97.3



gbs2018
SAL_2118
100.0
SAG2063
87.5
SAJ_1966
87.5

















TABLE 5







Sequence identity of proteins as determined by the first run of sequencing. % Id, amino


acid sequence identity of the respective protein in percentage as determined by DNA sequencing.


All comparisons are performed to the respective protein of GBS strain NEM316.














S. agalactiae
S. agalactiae
S. agalactiae
S. agalactiae
S. agalactiae
S. agalactiae


ORF name
IC105
IC458
12401
BAA23
COH1
0176H4A


(NEM316)
% Id
% Id
% Id
% Id
% Id
% Id
















gbs0233
99.0
100
99.0
99.0
99.7
99.7


gbs1087
86.0
95.2
93.9
91.5
92.7
98.6


gbs1309
100
100
99.3
100
99.5
99.5


gbs1477
48.9
49.3
48.8
98.4
Not
67.8







determined


gbs1478
88.2
88.1
87.7
99.1
Not
Not







determined
determined


gbs2018
77.6
99.7
77.2
78.5
Not
80.2







determined
















TABLE 6







Sequence identity of gbs1477 proteins as determined by the first run of


sequencing. % Id, amino acid sequence identity of the respective protein


in percentage as determined by DNA sequencing. Pairwise comparisons are


performed for the gbs1477 protein among the sequenced GBS strains.













S. agalactiae
S. agalactiae
S. agalactiae
S. agalactiae
S. agalactiae



IC105
IC458
12401
BAA23
0176H4A


gbs1477
% Id
% Id
% Id
% Id
% Id















S. agalactiae NEM316
48.9
49.3
48.8
98.4
67.8


S. agalactiae IC105
100
58.6
99.9
49.4
48.0


S. agalactiae IC458

100
58.6
49.5
53.2


S. agalactiae 12401


100
49.4
47.9


S. agalactiae BAA23



100
68.5


S. agalactiae 0176H4A




100
















TABLE 7





Amino acid and encoding DNA sequences of gbs0233 proteins derived from


different strains of S. agalactiae







Strain 0176H4A


ORF DNA sequence (SEQ ID NO: 61)


ATGCTTAAAAAATCGCACTTTTTACAGATATTTACACTTTGCTTAGCCCTCTTAACGATTTCTGGTTGTCAA


TTAACCGATACTAAAAAACCTGGTCATACCACAATTAAGGTTGCTGCACAAAGTTCTACAGAGTCTAGTATC


ATGGCAAATATTGTCACCGAATTAATTCATCACGAATTAGGATACAACACAACTTTAATAAGCAATCTTGGT


TCCTCTACGGTTACTCACCAAGCTTTGCTCCGTGGTGATGCTGACATTGCTGCCACACGTTATACAGGAACA


GACATCACAGGAACTCTTGGCTTAAAAGCTGTTAAAGACCCTAAAGAAGCTTCTAAGATTGTAAAAACTGAA


TTCCAAAAACGCTACAATCAAACTTGGTATCCTACTTATGGTTTTTCTGATACTTATGCATTCATGGTTACT


AAAGAGTTTGCCAGACAGAATAAAATCACCAAGATCTCTGATCTCAAAAAATTATCAACAACTATGAAGGCA


GGGGTTGATAGTTCATGGATGAATCGCGAGGGAGATGGATACACTGATTTCGCTAAAACATACGGTTTTGAA


TTTTCACATATTTACCCTATGCAAATTGGCTTAGTCTATGATGCGGTTGAAAGTAACAAAATGCAATCTGTA


TTAGGCTACTCCACTGACGGTCGTATTTCGAGCTATGATTTAGAAATTTTAAGGGATGATAAAAAATTCTTT


CCTCCTTATGAAGCCTCTATGGTTGTCAACAATTCTATCATCAAAAAAGATCCTAAACTAAAAAAATTACTC


CATCGACTCGATGGTAAAATCAATTTAAAAACGATGCAAAACCTTAATTATATGGTAGATGATAAACTTTTA


GAACCTTCAGTTGTTGCCAAACAATTTTTAGAAAAAAACCATTATTTTAGAGGAGATAAATAA


 


ORF amino acid sequence (SEQ ID NO: 55):


MLKKSHFLQIFTLCLALLTISGCQLTDTKKPGHTTIKVAAQSSTESSIMANIVTELIHHELGYNTTLISNLG


SSTVTHQALLRGDADIAATRYTGTDITGTLGLKAVKDPKEASKIVKTEFQKRYNQTWYPTYGFSDTYAFMVT


KEFARQNKITKISDLKKLSTTMKAGVDSSWMNREGDGYTDFAKTYGFEFSHIYPMQIGLVYDAVESNKMQSV


LGYSTDGRISSYDLEILRDDKKFFPPYEASMVVNNSIIKKDPKLKKLLHRLDGKINLKTMQNLNYMVDDKLL


EPSVVAKQFLEKNHYFRGDK


 


Strain 12401


ORF DNA sequence (SEQ ID NO: 62):


ATGCTTAAAAAATCGCACTTTTTACAGATATTTACACTTTGCTTAGCCCTCTTAACGATTTCTGGTTGTCAA


TTAACCGATACTAAAAAGTCTGGTCATACCACAATTAAGGTTGCTGCCCAAAGTTCTACAGAGTCTAGTATT


ATGGCAAATATTATCACCGAATTAATTCATCACGAATTAGGATACAACACAACTTTAATAAGCAATCTTGGT


TCCTCTACGGTTACTCACCAAGCTTTGCTCCGTGGTGATGCTGACATTGCTGCCACACGTTATACAGGAACA


GACATCACAGGGACTCTTGGTTTAAAAGCTGTTAAAGACCCTAAAGAAGCTTCTAAGATTGTAAAAACTGAA


TTCCAAAAACGCTACAATCAAACTTGGTATCCTACTTATGGTTTTTCTGATACTTATGCATTCATGGTTACT


AAAGAGTTTGCCAGACAgAATAAAATCACTAAGATCTCTGATCTTAAAAAATTATCAACAACTATGAAGGCA


GGGGTTGATAGTTCATGGATGAATCGCGAGGGAGATGGATACACTGATTTCGCTAAAACATACGGTTTTGAA


TTTTCACATATTTACCCTATGCAAATTGGCTTAGTCTATGATGCGGTTGAAAGTAACAAAATGCAATCTGTA


TTAGGCTACTCCACTGACGGTCGTATTTCGAGCTATGATTTAGAAATTTTAAGGGATGATAAAAAATTCTTT


CCTCCTTATGAAGCCTCTATGGTTGTCAACAATTCTATCATCAAAAAAGATCCTAAACTAAAAAAATTACTC


CATCGACTCGATGGTAAAATCAATTTAAAAACGATGCAAAACCTTAATTATATGGTAGATGATAAACTTTTA


GAACCTTCAGTTGTTGCCAAACAATTTTTAGAAAAAAACCATTATTTTAGAGGAGATAAATAA


 


ORF amino acid sequence (SEQ ID NO: 56):


MLKKSHFLQIFTLCLALLTISGCQLTDTKKSGHTTIKVAAQSSTESSIMANIITELIHHELGYNTTLISNLG


SSTVTHQALLRGDADIAATRYTGTDITGTLGLKAVKDPKEASKIVKTEFQKRYNQTWYPTYGFSDTYAFMVT


KEFARQNKITKISDLKKLSTTMKAGVDSSWMNREGDGYTDFAKTYGFEFSHIYPMQIGLVYDAVESNKMQSV


LGYSTDGRISSYDLEILRDDKKFFPPYEASMVVNNSIIKKDPKLKKLLHRLDGKINLKTMQNLNYMVDDKLL


EPSVVAKQFLEKNHYFRGDK


 


Strain BAA23


ORF DNA sequence (SEQ ID NO: 63):


ATGCTTAAAAAATCGCACTTTTTACAGATATTTACACTTTGCTTAGCCCTCTTAACGATTTCTGGTTGTCAA


TTAACCGATACTAAAAAGTCTGGTCATACCACAATTAAGGTTGCTGCCCAAAGTTCTACAGAGTCTAGTATT


ATGGCAAATATTATCACCGAATTAATTCATCACGAATTAGGATACAACACAACTTTAATAAGCAATCTTGGT


TCCTCTACGGTTACTCACCAAGCTTTGCTCCGTGGTGATGCTGACATTGCTGCCACACGTTATACAGGAACA


GACATCACAGGGACTCTTGGTTTAAAAGCTGTTAAAGACCCTAAAGAAGCTTCTAAGATTGTAAAAACTGAA


TTCCAAAAACGCTACAATCAAACTTGGTATCCTACTTATGGTTTTTCTGATACTTATGCATTCATGGTTACT


AAAGAGTTTGCCAGACAGAATAAAATCACTAAGATCTCTGATCTTAAAAAATTATCAACAACTATGAAGGCA


GGGGTTGATAGTTCATGGATGAATCGCGAGGGAGATGGATACACTGATTTCGCTAAAACATACGGTTTTGAA


TTTTCACATATTTACCCTATGCAAATTGGCTTAGTCTATGATGCGGTTGAAAGTAACAAAATGCAATCTGTA


TTAGGCTACTCCACTGACGGTCGTATTTCGAGCTATGATTTAGAAATTTTAAgGGATGATAAAAAATTCTTT


CCTCCTTATGAAGCCTCTATGGTTGTCAACAATTCTATCATCAAAAAAGATCCTAAACTAAAAAAATTACTC


CATCGACTCGATGGTAAAATCAATTTAAAAACGATGCAAAACCTTAATTATATGGTAGATGATAAACTTTTA


GAACCTTCAGTTGTTGCCAAACAATTTTTAGAAAAAAACCATTATTTTAGAGGAGATAAATAA


 


ORF amino acid sequence (SEQ ID NO: 57):


MLKKSHFLQIFTLCLALLTISGCQLTDTKKSGHTTIKVAAQSSTESSIMANIITELIHHELGYNTTLISNLG


SSTVTHQALLRGDADIAATRYTGTDITGTLGLKAVKDPKEASKIVKTEFQKRYNQTWYPTYGFSDTYAFMVT


KEFARQNKITKISDLKKLSTTMKAGVDSSWMNREGDGYTDFAKTYGFEFSHIYPMQIGLVYDAVESNKMQSV


LGYSTDGRISSYDLEILRDDKKFFPPYEASMVVNNSIIKKDPKLKKLLHRLDGKINLKTMQNLNYMVDDKLL


EPSVVAKQFLEKNHYFRGDK


 


Strain COH1


ORF DNA sequence (SEQ ID NO: 64):


ATGCTTAAAAAATCGCACTTTTTACAGATATTTACACTTTGCTTAGCCCTCTTAACGATTTCTGGTTGTCAA


TTAACCGATACTAAAAAACCTGGTCATACCACAATTAAGGTTGCTGCCCAAAGTTCTACAGAGTCTAGTATC


ATGGCAAATATTGTCACCGAATTAATTCATCACGAATTAGGATACAACACAACTTTAATAAGCAATCTTGGT


TCCTCTACGGTTACTCACCAAGCTTTGCTCCGTGGTGATGCTGACATTGCTGCCACACGTTATACAGGAACA


GACATCACAGGAACTCTTGGCTTAAAAGCTGTTAAAGACCCTAAAGAAGCTTCTAAGATTGTAAAAACTGAA


TTCCAAAAACGCTACAATCAAACTTGGTATCCTACTTATGGTTTTTCTGATACTTATGCATTCATGGTTACT


AAAGAGTTTGCCAGACAGAATAAAATCACCAAGATCTCTGATCTCAAAAAGTTATCAACAACTATGAAGGCA


GGGGTTGATAGTTCATGGATGAATCGCGAGGGAGATGGATACACTGATTTCGCTAAAACATACGGTTTTGAA


TTTTCACATATTTACCCTATGCAAATTGGCTTAGTCTATGATGCGGTTGAAAGTAACAAAATGCAATCTGTA


TTAGGCTACTCCACTGACGGTCGTATTTCGAGCTATGATTTAGAAATTTTAAGGGATGATAAAAAATTCTTT


CCTCCTTATGAAGCCTCTATGGTTGTCAACAATTCTATCATCAAAAAAGATCCTAAACTAAAAAAATTACTC


CATCGACTCGATGGTAAAATCAATTTAAAAACGATGCAAAACCTTAATTATATGGTAGATGATAAACTTTTA


GAACCTTCAGTTGTTGCCAAACAATTTTTAGAAAAAAACCATTATTTTAGAGGAGATAAATAA


 


ORF amino acid sequence (SEQ ID NO: 58):


MLKKSHFLQIFTLCLALLTISGCQLTDTKKPGHTTIKVAAQSSTESSIMANIVTELIHHELGYNTTLISNLG


SSTVTHQALLRGDADIAATRYTGTDITGTLGLKAVKDPKEASKIVKTEFQKRYNQTWYPTYGFSDTYAFMVT


KEFARQNKITKISDLKKLSTTMKAGVDSSWMNREGDGYTDFAKTYGFEFSHIYPMQIGLVYDAVESNKMQSV


LGYSTDGRISSYDLEILRDDKKFFPPYEASMVVNNSIIKKDPKLKKLLHRLDGKINLKTMQNLNYMVDDKLL


EPSVVAKQFLEKNHYFRGDK


 


Strain IC105


ORF DNA sequence (SEQ ID NO: 65):


ATGCTTAAAAAATCGCACTTTTTACAGATATTTACACTTTGCTTAGCCCTCTTAACGATTTCTGGTTGTCAA


TTAACCGATACTAAAAAGTCTGGTCATACCACAATTAAGGTTGCTGCCCAAAGTTCTACAGAGTCTAGTATT


ATGGCAAATATTATCACCGAATTAATTCATCACGAATTAGGATACAACACAACTTTAATAAGCAATCTTGGT


TCCTCTACGGTTACTCACCAAGCTTTGCTCCGTGGTGATGCTGACATTGCTGCCACACGTTATACAGGAACA


GACATCACAGGGACTCTTGGTTTAAAAGCTGTTAAAGACCCTAAAGAAGCTTCTAAGATTGTAAAAACTGAA


TTCCAAAAACGCTACAATCAAACTTGGTATCCTACTTATGGTTTTTCTGATACTTATGCATTCATGGTTACT


AAAGAGTTTGCCAGACAGAATAAAATCACTAAGATCTCTGATCTTAAAAAATTATCAACAACTATGAAGGCA


GGGGTTGATAGTTCATGGATGAATCGCGAGGGAGATGGATACACTGATTTCGCTAAAACATACGGTTTTGAA


TTTTCACATATTTACCCTATGCAAATTGGCTTAGTCTATGATGCGGTTGAAAGTAACAAAATGCAATCTGTA


TTAGGCTACTCCACTGACGGTCGTATTTCGAGCTATGATTTAGAAATTTTAAGGGATGATAAAAAATTCTTT


CCTCCTTATGAAGCCTCTATGGTTGTCAACAATTCTATCATCAAAAAAGATCCTAAACTAAAAAAATTACTC


CATCGACTCGATGGTAAAATCAATTTAAAAACGATGCAAAACCTTAATTATATGGTAGATGATAAACTTTTA


GAACCTTCAGTTGTTGCCAAACAATTTTTAGAAAAAAACCATTATTTTAGAGGAGATAAATAA


 


ORF amino acid sequence (SEQ ID NO: 59):


MLKKSHFLQIFTLCLALLTISGCQLTDTKKSGHTTIKVAAQSSTESSIMANIITELIHHELGYNTTLISNLG


SSTVTHQALLRGDADIAATRYTGTDITGTLGLKAVKDPKEASKIVKTEFQKRYNQTWYPTYGFSDTYAFMVT


KEFARQNKITKISDLKKLSTTMKAGVDSSWMNREGDGYTDFAKTYGFEFSHIYPMQIGLVYDAVESNKMQSV


LGYSTDGRISSYDLEILRDDKKFFPPYEASMVVNNSIIKKDPKLKKLLHRLDGKINLKTMQNLNYMVDDKLL


EPSVVAKQFLEKNHYFRGDK


 


Strain IC458


ORF DNA sequence (SEQ ID NO: 66):


ATGCTTAAAAAATCGCACTTTTTACAGATATTTACACTTTGCTTAGCCCTCTTAACGATTTCTGGTTGTCAA


TTAACCGATACTAAAAAACCTGGTCATACCACAATTAAGGTTGCTGCCCAAAGTTCTACAGAGTCTAGTATC


ATGGCAAATATTGTCACCGAATTAATTCATCACGAATTAgGATACAACACAACTTTAATAAGCAATCTTGGT


TCCTCTACGGTTACTCACCAAGCTTTGCTCCGTGGTGATGCTGACATTGCTGCCACACGTTATACAGGAACA


GACATCAcAGGAACTCTTGGCTTAAAAGCTGTTAAAGACACTAAAGAAGCTTCTAAGATTGTAAAAACTGAA


TTCCAAAAACGCTACAATCAAACTTGGTATCCTACTTATGGTTTTTCTGATACTTATGCATTCATGGTTACT


AAAgAGTTTGCCAGACAGAAtAAAATCACCAAGATCTCTGAtCTCAAAAAgTTATCAACAACTATGAAGGCA


GGGGTTGATAGTTCATGGATGAATCGCGAGGGAGATGGATACACTGATTTCGCTAAAACATACGGTTTTGAA


TTTTCACATATTTACCCTATGCAAATTGGCTTAGTCTATGATGCAGTTGAAAGTAACAAAATGCAATCTGTA


TTAGGCTACTCCACTGACGGTCGTATTTCGAGCTATGATTTAGAAATTTTAAGGGATGATAAAAAATTCTTT


CCTCCTTATGAAGCCTCTATGGTTGTCAACAATTCTATCATCAAAAAAGATCCTAAACTAAAAAAATTACTC


CATCGACTCGATGGTAAAATCAATTTAAAAACGATGCAAAACCTTAATTATATGGTAGATGATAAACTTTTA


GAACCTTCAGTTGTTGCCAAACAATTTTTAGAAAAAAACCATTATTTTAGAGGAGATAAATAA


 


ORF amino acid sequence (SEQ ID NO: 60):


MLKKSHFLQIFTLCLALLTISGCQLTDTKKPGHTTIKVAAQSSTESSIMANIVTELIHHELGYNTTLISNLG


SSTVTHQALLRGDADIAATRYTGTDITGTLGLKAVKDTKEASKIVKTEFQKRYNQTWYPTYGFSDTYAFMVT


KEFARQNKITKISDLKKLSTTMKAGVDSSWMNREGDGYTDFAKTYGFEFSHIYPMQIGLVYDAVESNKMQSV


LGYSTDGRISSYDLEILRDDKKFFPPYEASMVVNNSIIKKDPKLKKLLHRLDGKINLKTMQNLNYMVDDKLL


EPSVVAKQFLEKNHYFRGDK
















TABLE 8





Amino acid and encoding DNA sequences of gbs1087 proteins derived from


different strains of S. agalactiae







Strain 0176H4A


ORF DNA sequence (SEQ ID NO: 73):


TTGTTCAATAAAATAGGTTTTAGAACTTGGAAATCAGGAAAGCTTTGGCTTTATATGGGAGTGCTAGGATCA


ACTATTATTTTAGGATCAAGTCCTGTATCTGCTATGGATAGTGTTGGAAATCAAAGTCAGGGCAATGTTTTA


GAGCGTCGTCAACGCGATGCAGAAAACAGAAGCCAAGGTAATGTTCTAGAGCGTCGTCAACGCGATGCAGAA


AACAGAAGCCAAGGTAATGTTCTAGAGCGTCGTCAACGTGATGCGGAAAACAAGAGCCAAGTAGGTCAACTT


ATAGGGAAAAATCCACTTCTTTCAAAGTCAATTATATCTAGAGAAAATAATCACTCTAGTCAAGGTGACTCT


AACAAACAGTCATTCTCTAAAAAAGTATCTCAGGTTACTAATGTAGCTAATAGACCGATGTTAACTAATAAT


TCTAGAACAATTTCAGTGATAAATAAATTACCTAAAACAGGTGATGATCAAAATGTCATTTTTAAACTTGTA


GGTTTTGGTTTAATTTTGTTAACAAGTCGCTGCGGTTTGAGACGCAATGAAAATTAA


 


ORF amino acid sequence (SEQ ID NO: 67):


LFNKIGFRTWKSGKLWLYMGVLGSTIILGSSPVSAMDSVGNQSQGNVLERRQRDAENRSQGNVLERRQRDAE


NRSQGNVLERRQRDAENKSQVGQLIGKNPLLSKSIISRENNHSSQGDSNKQSFSKKVSQVTNVANRPMLTNN


SRTISVINKLPKTGDDQNVIFKLVGFGLILLTSRCGLRRNEN


 


Strain 12401


ORF DNA sequence (SEQ ID NO: 74):


TTGTTCAATAAAATAGGTTTTAGAACTTGGAAATCAGGAAAGCTTTGGCTTTATATGGGAGTGCTAGGATCA


ACTATTATTTTAGGATCAAGTCCTGTATCTGCTATGGATAGTGTTGGAAATCAAAGTCAAGGTAATGTTCTA


GAGCGTCGTCAACGTGATGCGGATAACAAGAGCCAAGGTAATGTTCTAGAACGTCGTCAACGCGATGTGGAA


AACAAAAGTCAGGGCAATGTTCTAGAACGTCGTCAACGTGATGTTGAGAATAAGAGCCAAGGCAATGTTCTA


GAGCGTCGCCAACGTGATGCAGAAAACAAAAGTCAGGGTAATGTTCTAGAGCGTCGTCAACGCGATGCAGAT


AACAAGAGCCAAGGTAATGTTCTAGAACGTCGTCAACGCGATGTGGAAAACAAAAGTCAGGGCAATGTTCTA


GAGCGTCGCCAACGTGATGTTGAGAACAAGAGCCAAGTAGGTCAACTTATAGGGAAAAATCCACTTCTTTCA


AAGTCAACTATATCTAGAGAAAATAATCACTCTAGTCAAGGTGACTCTAACAAACAGTCATTCTCTAAAAAA


GTATCTCAGGTTACTAATGTAGCTAATAGACCAATGTTAACTAATAATTCTAGAACAATTTCAGTGATAAAT


AAATTACCTAAAACAGGTGATGATCAAAATGTCATTTTTAAACTTGTAGGTTTTGGTTTAATTTTGTTAACA


AGTCGCTGCGGTTTGAGACGCAATGAAAATTAA


 


ORF amino acid sequence (SEQ ID NO: 68):


LFNKIGFRTWKSGKLWLYMGVLGSTIILGSSPVSAMDSVGNQSQGNVLERRQRDADNKSQGNVLERRQRDVE


NKSQGNVLERRQRDVENKSQGNVLERRQRDAENKSQGNVLERRQRDADNKSQGNVLERRQRDVENKSQGNVL


ERRQRDVENKSQVGQLIGKNPLLSKSTISRENNHSSQGDSNKQSFSKKVSQVTNVANRPMLTNNSRTISVIN


KLPKTGDDQNVIFKLVGFGLILLTSRCGLRRNEN


 


Strain BAA23


ORF DNA sequence (SEQ ID NO: 75):


TTGTTCAATAAAATAGGTTTTAGAACTTGGAAATCAGGAAAGCTTTGGCTTTATATGGGAGTGCTAGGATCA


ACTATTATTTTAGGATCAAGTCCTGTATCTGCTATGGATAGTGTTGGAAATCAAAGTCAGGGCAATGTTTTA


GAGCGTCGTCAACGTGATGCAGAAAACAGAAGCCAAGGTAATGTTCTAGAGCGTCGTCAACGTGATGCAGAA


AACAGAAGCCAAGGTAATGTTCTAGAGCGTCGTCAACGCGATGCAGAAAACAGAAGCCAAGGTAATGTTCTA


GAGCGTCGTCAACGTGATGTTGAGAATAAGAGCCAAGGCAATGTTTTAGAGCGTCGTCAACGCGATGTTGAG


AATAAGAGCCAAGGTAATGTTCTAGAGCGTCGTCAACGTGATGCGGAAAACAAGAGCCAAGGCAATGTTCTA


GAGCGTCGTCAACGTGATGCGGAAAACAAGAGCCAAGGCAATGTTCTAGAGCGTCGTCAACGCGATGCAGAA


AACAGAAGCCAAGGTAATGTTTTAGAACGTCGTCAACGCGATGTTGAGAACAAGAGCCAAGGTAACGTTCTA


GAGCGTCGCCAACGTGACGTTGAGAACAAGAGCCAAGGTAATGTTTTAGAGCGTCGCCAACGCGATGCGGAT


AACAAAAGTCAGGGCAATGTTTTAGAGCGTCGCCAACGTGATGTTGAGAACAAGAGCCAAGGTAATGTTCTA


GAGCGTCGCCAAAATAATGTCCTTATTAAGAGTCAAGATAATGTTCTAGAGCGCCGCCAACGTGATGCGGAT


AACAAGAGCCAGGGTAACGTTCTAGAGCGTCGTCAACGCGATGTTGAGAATAAGAGCCAAGGTAATGTTTTA


GAGCGTCGCCAACATGATGTTGAGAATAAGAGTCAAGTAGGTCAACTTATAGGGAAAAATCCACTTTTTTCA


AAGTCAACTGTATCTAGAGAAAATAATCACTCTAGTCAAGGTGACTCTAACAAACAGTCATTCTCTAAAAAA


GTATCTCAGGTTACTAATGTAGCTAATAGACCGATGTTAACTAATAATTCTAGAACAATTTCAGTGATAAAT


AAATTACCTAAAACAGGTGATGATCAAAATGTCATTTTTAAACTTGTAGGTTTTGGTTTAATTTTATTAACA


AGTCTCTGCGGTTTGAGACGCAATGAAAATTAA


 


ORF amino acid sequence (SEQ ID NO: 69):


LFNKIGFRTWKSGKLWLYMGVLGSTIILGSSPVSAMDSVGNQSQGNVLERRQRDAENRSQGNVLERRQRDAE


NRSQGNVLERRQRDAENRSQGNVLERRQRDVENKSQGNVLERRQRDVENKSQGNVLERRQRDAENKSQGNVL


ERRQRDAENKSQGNVLERRQRDAENRSQGNVLERRQRDVENKSQGNVLERRQRDVENKSQGNVLERRQRDAD


NKSQGNVLERRQRDVENKSQGNVLERRQNNVLIKSQDNVLERRQRDADNKSQGNVLERRQRDVENKSQGNVL


ERRQHDVENKSQVGQLIGKNPLFSKSTVSRENNHSSQGDSNKQSFSKKVSQVTNVANRPMLTNNSRTISVIN


KLPKTGDDQNVIFKLVGFGLILLTSLCGLRRNEN


 


Strain COH1


ORF DNA sequence (SEQ ID NO: 76):


TTGTTCAATAAAATAGGTTTTAGAACTTGGAAATCAGGAAAGCTTTGGCTTTATATGGGAGTGCTAGGATCA


ACTATTATTTTAGGATCAAGTCCTGTATCTGCTATGGATAGTGTTGGAAATCAAAGTCAAGGTAATGTTTTA


GAGCGTCGCCAACGTGATGCGGAAAACAAAAGTCAGGGTAATGTTTTAGAGCGTCGCCAACGTGATGCGGAA


AACAAGAGCCAAGGCAATGTTTTAGAGCGTCGTCAACGCGATGTTGAGAATAAGAGCCAAGGCAATGTTTTA


GAGCGTCGTCAACGTGATGCGGAAAACAAAAGTCAGGGCAATGTTCTAGAGCGCCGCCAACGTGATGCGGAT


AACAAGAGCCAAGTAGGTCAACTTATAGGGAAAAATCCACTTTTTTCAAAGCCAACTGTATCTAGAGAAAAT


AATCACTCTAGTCAAGGTGACTCTAACAAACAGTCATTCTCTAAAAAAGTATCTCAGGTTACTAATGTAGCT


AATAGACCGATGTTAACTAATAATTCTAGAACAATTTCAGTGATAAATAAATTACCTAAAACAGGTGGTGAT


CAAAATGTCATTTTTAAACTTGTAGGTTTTGGTTTAATTTTGTTAACAAGTCGCTGCGGTTTGAGACGCAAT


GAAAATTAA


 


ORF amino acid sequence (SEQ ID NO: 70):


LFNKIGFRTWKSGKLWLYMGVLGSTIILGSSPVSAMDSVGNQSQGNVLERRQRDAENKSQGNVLERRQRDAE


NKSQGNVLERRQRDVENKSQGNVLERRQRDAENKSQGNVLERRQRDADNKSQVGQLIGKNPLFSKPTVSREN


NHSSQGDSNKQSFSKKVSQVTNVANRPMLTNNSRTISVINKLPKTGGDQNVIFKLVGFGLILLTSRCGLRRN


EN


 


Strain IC105


ORF DNA sequence (SEQ ID NO: 77):


TTGTTCAATAAAATAGGTTTTAGAACTTGGAAATCAGGAAAGCTTTGGCTTTATATGGGAGTGCTAGGATCa


actattattttaggatcaagtcctgtttctgCTATGGATAGTGATGGAAATCAAAGTCAGGGCAATGTTTTA


GAGCGTCGTCAACGTGATGCGGAAAACAAGAGCCAAGGCAATGTTCTAGAGCGTCGTCAACGCGATGCAGAA


AACAGAAGCCAAGGTAATGATCTAGAGCGTCGCCAACGTGACGTTGAGAACAAGAGCCAAGGTAACGTTCTA


GAGCGTCGTCAACGTGATGCAGATAACAAGAGCCAGGGCAATGTTTTAGAGCGTCGCCAACGTGATGTTGAG


AACAAGAGCCAAGGTAATGTTCTAGAGCGTCGCCAAAATAATGTCCTTATTAAGAGTCAAGATAATGATCTA


GAGCGCCGCCAACGTGATGCGGATAACAAGAGCCAGGGTAACGTTCTAGAGCGTCGCCAACGTGATGTTGAG


AACAAGAGCCAAGGTAATGTTCTAGAGCGCCGCCAACGTGATGCGGATAACAAGAGCCAGGGTAACGTTCTA


GAGTGTCGCCAACGTGATGTTGAGAACAAGAGCCAAGGTAATGTTCTAGAGCGTCGTCAACGTGATGCGGAA


AACAAGAGCCAAGGCAATGTTCTAGAGCGTCGTCAACGCGATGCAGAAAACAGAAGCCAAGGTAATGTTCTA


GAGCGTCGCCAACGTGACGTTGAGAACAAGAGCCAAGGTAACGTTCTAGAGCGTCGTCAACGTGATGCAGAT


AACAAGAGCCAGGGCAATGTTTTAGAGCGTCGCCAACGTGATGTTGAGAACAAGAGCCAAGGTAATGTTCTA


GAGCGTCGCCAAAATAATGTCCTTATTAAGAGTCAAGATAATGTTCTAGAGCGCCGCCAACGTGATGCGGAT


AACAAGAGCCAGGGTAACGTTCTAGAGCGTCGCCAACGTGATGTTGAGAACAAGAGCCAAGGCAATGTTTTA


GAGCGTCGTCAACGCGATGTTGAGAATAAGAGTCAAGTAGGTCAACTTATAGGGAAAAATCCACTTTTTTCA


AAGTCAACTGTATCTAGAGAAAATAATCACTCTAGTCAAGGTGACTCTAACAAACAGTCATTCTCTAAAAAA


GTATCTCAGGTTACTAATGTAGCTAATAGACCGATGTTAACTAATAATTCTAGAACAATTTCAGTGATAAAT


AAATTACCTAAAACAGGTGATGATCAAAATGTCATTTTTAAACTTGTAGGTTTTGGTTTAATTTTATTAACA


AGTCTCTGCGGTTTGAGACGCAATGAAAATTAA


 


ORF amino acid sequence (SEQ ID NO: 71):


LFNKIGFRTWKSGKLWLYMGVLGSTIILGSSPVSAMDSDGNQSQGNVLERRQRDAENKSQGNVLERRQRDAE


NRSQGNDLERRQRDVENKSQGNVLERRQRDADNKSQGNVLERRQRDVENKSQGNVLERRQNNVLIKSQDNDL


ERRQRDADNKSQGNVLERRQRDVENKSQGNVLERRQRDADNKSQGNVLECRQRDVENKSQGNVLERRQRDAE


NKSQGNVLERRQRDAENRSQGNVLERRQRDVENKSQGNVLERRQRDADNKSQGNVLERRQRDVENKSQGNVL


ERRQNNVLIKSQDNVLERRQRDADNKSQGNVLERRQRDVENKSQGNVLERRQRDVENKSQVGQLIGKNPLFS


KSTVSRENNHSSQGDSNKQSFSKKVSQVTNVANRPMLTNNSRTISVINKLPKTGDDQNVIFKLVGFGLILLT


SLCGLRRNEN


 


Strain IC458


ORF DNA sequence (SEQ ID NO: 78):


TTGTTCAATAAAATAGGTTTTAGAACTTGGAAATCAGGAAAGCTTTGGCTTTATATGGGAGTGCTAGGATCA


ACTATTATTTTAGGATCAAGTCCTGTATCTGCTATGGATAGTGTTGGAAATCAAAGTCAGGGCAATGTTTTA


GAGCGTCGTCAACGCGATGTTGAGAATAAGAGCCAAGGTAATGTTCTAGAGCGTCGTCAACGTGATGCGGAA


AACAAGAGCCAAGGCAATGTTTTAGAGCGTCGTCAACGTGATGCAGAAAACAGAAGCCAAGGCAATGTTTTA


GAGCGTCGTCAACGTGATGCAGAAAACAGAAGCCAAGGCAATGTTTTAGAGCGTCGTCAACGCGATGTTGAG


AATAAGAGCCAAGGTAATGTTCTAGAGCGTCGTCAACGCGATGTTGAGAATAAGAGCCAAGGTAATGTTCTA


GAGCGTCGTCAACGTGATGCGGAAAACAAGAGCCAAGGCAATGTTTTAGAGCGTCGTCAACGTGATGCAGAA


AACAGAAGCCAAGGCAATGTTTTAGAGCGTCGTCAACGTGATGCAGAAAACAGAAGCCAAGGCAATGTTTTA


GAGCGTCGTCAACGCGATGTTGAGAATAAGAGCCAAGGCAATGTTTTAGAGCGTCGTCAACGTGATGCAGAA


AACAGAAGCCAAGGCAATGTTTTAGAGCGTCGTCAACGTGATGCAGAAAACAGAAGCCAAGTAGGTCAACTT


ATAGGGAAAAATCCACTTCTTTCAAAGTCAATTATATCTAGAGAAAATAATCACTCTAGTCAAGGTGACTCT


AACAAACAGTCATTCTCTAAAAAAGTATCTCAGGTTACTAATGTAGCTAATAGACCGATGTTAACTAATAAT


TCTAGAACAATTTCAGTGATAAATAAATTACCTAAAACAGGTGATGATCAAAATGTCATTTTTAAACTTGTA


GGTTTTGGTTTAATTTTGTTAACAAGTCGCTGCGGTTTGAGACGCAATGAAAATTAA


 


ORF amino acid sequence (SEQ ID NO: 72):


LFNKIGFRTWKSGKLWLYMGVLGSTIILGSSPVSAMDSVGNQSQGNVLERRQRDVENKSQGNVLERRQRDAE


NKSQGNVLERRQRDAENRSQGNVLERRQRDAENRSQGNVLERRQRDVENKSQGNVLERRQRDVENKSQGNVL


ERRQRDAENKSQGNVLERRQRDAENRSQGNVLERRQRDAENRSQGNVLERRQRDVENKSQGNVLERRQRDAE


NRSQGNVLERRQRDAENRSQVGQLIGKNPLLSKSIISRENNHSSQGDSNKQSFSKKVSQVTNVANRPMLTNN


SRTISVINKLPKTGDDQNVIFKLVGFGLILLTSRCGLRRNEN
















TABLE 9





Amino acid and encoding DNA sequences of gbs1309 proteins derived from


different strains of S. agalactiae







Strain 0176H4A


ORF DNA sequence (SEQ ID NO: 85):


TTTAGTGTAACCTATTCACAGTCTGAACGTACGGTTGTTTTCTCTTTTGGAGAAATAACATTTAGTAGGAGT


CGCTGGACAAATGGCTTTGAAACTAGAATACCAGTAGATGAGTGGTTAGGTCTTGAAAAATATAAGAGATAT


TCAATAGAATTCTTATATCATGTTGCAAAATTGGCTACAATGATGCCTTATCGTCAAGTTTGCAAAGTAATA


GATAGCACTTTGCAAACAATCATAACAAAAGACTGTGTTTTAAAAGCAGTAAAATTTGTAGAAAAATTGTTA


AAAGAAAAAGAACGCTATCGTTTTTATTTGGAAGAGCCACCCGAACGTAAAAAAGTGAAAAAACTGTATGTT


GAGGGTGATGGAGTCATGATTAAAAGCACAGATTCTAGAGAGGAAAGAAGGTATTTAGATTTAACACATTTT


GTTATTCATACAGGCTCAAAAAAAGTTTCTACTAAAAGATATGAATTGCAGGACAAGCACGAAATATTACAG


CTTAATTATGATAAAGCTAAATATAATCTTTTAGATTATATTTATAATAACTATGAAGTAGATGACGATACT


ATTTTAATCACTAACTCTGATATGGGTAAAGGCTATACTAGTAGAGTTTTTAAGGAATTAGGAAAAGCACTT


AAGGTAAAGAAACATGAGCATTTTTGGGATATCTATCATGTTAAAGAAAAGTTAAGTTCATACCTTAGAAAA


TATCCAATTGAATTAACCGATTTTACTTTAGATGCGGTAAAAAAATATAATTCTGATAAGCTTGAATTAGTT


TTTGATACTGTTGAATCACTGATTTGTGATGAACTTGAAGATCAAGAATTTCAGAAGTTTAAGAAAAAAGTA


TTAAATAATTTCAAATATATAAAACCAGCTCATCTTAGAAATCTTTCAAATCGTGGTATTGGTATCATGGAA


TCACAACACAGAAAGATAACGTATAGAATGAAGCGACGTGGCATGTATTGGTCAAAGTGGGGAATCTCCACA


ATGGCAAATATGATTATATTTGAAAGAGCTAACGGTTTACGAGAATTATTTTTCGGTTCTTGGAGAAAGGTA


TACAGTGAGTATAAAGAAGGTTCATTTAGTGCAGGGCGACTTTTTAAAAAGACAGATGAATTAGATAAATTT


TCTAAGCCCCTTCTAAAAAATGGCAGAAAATGGAGTATAACAGGAATCAAAACAAAATAG


 


ORF amino acid sequence (SEQ ID NO: 79):


FSVTYSQSERTVVFSFGEITFSRSRWTNGFETRIPVDEWLGLEKYKRYSIEFLYHVAKLATMMPYRQVCKVI


DSTLQTIITKDCVLKAVKFVEKLLKEKERYRFYLEEPPERKKVKKLYVEGDGVMIKSTDSREERRYLDLTHF


VIHTGSKKVSTKRYELQDKHEILQLNYDKAKYNLLDYIYNNYEVDDDTILITNSDMGKGYTSRVFKELGKAL


KVKKHEHFWDIYHVKEKLSSYLRKYPIELTDFTLDAVKKYNSDKLELVFDTVESLICDELEDQEFQKFKKKV


LNNFKYIKPAHLRNLSNRGIGIMESQHRKITYRMKRRGMYWSKWGISTMANMIIFERANGLRELFFGSWRKV


YSEYKEGSFSAGRLFKKTDELDKFSKPLLKNGRKWSITGIKTK


 


Strain 12401


ORF DNA sequence (SEQ ID NO: 86):


TTTAGTGTAACCTATTCACAGTCTGAACGTACGGTTGTTTTCTCTTTTGGAGAAATAACATTTAGTAGGAGT


CGCTGGACAAATGGCTTTGAAACTAGAATACCAGTAGATGAGTGGTTAGGTCTTGAAAAATATAAGAGATAT


TCAATAGAATTCTTATATCATGTTGCAAAATTGGCTACAATGATGCCTTATtGTCAAGTTTGCAAAGTAATA


GATAGCACTTTGCAAACAATCATAACAAAAGACTGTGTTTTAAAAGCAGTAAAATTTGTAGAAAAATTGTTA


AAAGAAAAAGAACGCTATCGTTTTTATTTGGAAGAGCCACCCGAACgTAAAAAAGtGAAAAAACTGTATGTT


GaGGGTGATGGAGTCATGATTAAAAGCACAGATTCTAGAGAGGAAAGAAGGTATTTAGATTTAACACATTTT


GTTATTCATACAGGCTCAAAAAAAGTTTCTACTAAAAGATATGAATTGCAGGACAAGCACGAAATATTACAG


CTTAATTATGATAAAGCTAAATATAATCTTTTAGATTATATTTATAATAACTATGAAGTAGATGACGATACT


ATTTTAATCACTAACTCTGATATGGGTAAAGGCTATACTAGTAGAGTTTTTAAGGAATTAGGAAAAGCACTT


AAGGTAAAGAAACATGAGCATTTTTGGGATATCTATCATGTtAAAGAAAAGTTAAGTTCATACCTTAGAAAA


TATCCAATTGAATTAACCGATTTTGCTTTAGATGCGGTAAAAAAATATAATTCTGATAAGCTTGAATTAGTT


TTTGATACTGTTGAATCACTGATTTGTGATGAACTTGAAGATCAAGAATTTCAGAAGTTTAAGAAAAAAGTA


TTAAATAATTTCAAATATATAAAACCAGCTCATCTTAGAAATCTTTCAAATCGTGGTATTGGTATCATGGAA


TCACAACACAGAAAGATAACGTATAGAATGAAGCGACGTGGCATGTATTGGTCAAAGTGGGGAATCTCCACA


ATGGCAAATATGATTATATTTGAAAGAGCTAACGGTTTACGAGAATTATTTTTCGGTTCTTGGAGAAAGGTA


TACAGTGAGTATAAAGAAGGTTCATTTAGTGCAGGGCGACTTTTTAAAAAGACAGATGAATTATATAAATTT


TCTAAGCCCCTTCTAAAAAATGGCAGAAAATGGAGTATAACAGGAATCAAAACAAAATAG


 


ORF amino acid sequence (SEQ ID NO: 80):


FSVTYSQSERTVVFSFGEITFSRSRWTNGFETRIPVDEWLGLEKYKRYSIEFLYHVAKLATMMPYCQVCKVI


DSTLQTIITKDCVLKAVKFVEKLLKEKERYRFYLEEPPERKKVKKLYVEGDGVMIKSTDSREERRYLDLTHF


VIHTGSKKVSTKRYELQDKHEILQLNYDKAKYNLLDYIYNNYEVDDDTILITNSDMGKGYTSRVFKELGKAL


KVKKHEHFWDIYHVKEKLSSYLRKYPIELTDFALDAVKKYNSDKLELVFDTVESLICDELEDQEFQKFKKKV


LNNFKYIKPAHLRNLSNRGIGIMESQHRKITYRMKRRGMYWSKWGISTMANMIIFERANGLRELFFGSWRKV


YSEYKEGSFSAGRLFKKTDELYKFSKPLLKNGRKWSITGIKTK


 


Strain BAA23


ORF DNA sequence (SEQ ID NO: 87):


TTTAGTGTAACCTATTCACAGTCTGAACGTACGGTTGTTTTCTCTTTTGGAGAAATAACATTTAGTAGGAGT


CGCTGGACAAATGGCTTTGAAACTAGAATACCAGTAGATGAGTGGTTAGGTCTTGAAAAATATAAGAGATAT


TCAATAGAATTCTTATATCATGTTGCAAAATTGGCTACAATGATGCCTTATCGTCAAGTTTGCAAAGTAATA


GATAGCACTTTGCAAACAATCATAACAAAAGACTGTGTTTTAAAAGCAGTAAAATTTGTAGAAAAATTGTTA


AAAGAAAAAGAACGCTATCGTTTTTATTTGGAAGAGCCACCCGAACGTAAAAAAGTGAAAAAACTGTATGTT


GAGGGTGATGGAGTCATGATTAAAAGCACAGATTCTAGAGAGGAAAGAAGGTATTTAGATTTAACACATTTT


GTTATTCATACAGGCTCAAAAAAAGTTTCTACTAAAAGATATGAATTGCAGGACAAGCACGAAATATTACAG


CTTAATTATGATAAAGCTAAATATAATCTTTTAGATTATATTTATAATAACTATGAAGTAGATGACGATACT


ATTTTAATCACTAACTCTGATATGGGTAAAGGCTATACTAGTAGAGTTTTTAAGGAATTAGGAAAAGCACTT


AAGGTAAAGAAACATGAGCATTTTTGGGATATCTATCATGTTAAAGAAAAGTTAAGTTCATACCTTAGAAAA


TATCCAATTGAATTAACCGATTTTGCTTTAGATGCGGTAAAAAAATATAATTCTGATAAGCTTGAATTAGTT


TTTGATACTGTTGAATCACTGATTTGTGATGAACTTGAAGATCAAGAATTTCAGAAGTTTAAGAAAAAAGTA


TTAAATAATTTCAAATATATAAAACCAGCTCATCTTAGAAATCTTTCAAATCGTGGTATTGGTATCATGGAA


TCACAACACAGAAAGATAACGTATAGAATGAAGCGACGTGGCATGTATTGGTCAAAGTGGGGAATCTCCACA


ATGGCAAATATGATTATACTTGAAAGAGCTAACGGTTTACGAGAATTATTTTTCGGTTCTTGGAGAAAGGTA


TACAGTGAGTATAAAGAAGGTTCATTTAGTGCAGGGCGACTTTTTAAAAAGACAGATGAATTAGATAAATTT


TCTAAGCCCCTTCTAAAAAATGGCAGAAAATGGAGTATAACAGGAATCAAAACAAAATAG


 


ORF amino acid sequence (SEQ ID NO: 81):


FSVTYSQSERTVVFSFGEITFSRSRWTNGFETRIPVDEWLGLEKYKRYSIEFLYHVAKLATMMPYRQVCKVI


DSTLQTIITKDCVLKAVKFVEKLLKEKERYRFYLEEPPERKKVKKLYVEGDGVMIKSTDSREERRYLDLTHF


VIHTGSKKVSTKRYELQDKHEILQLNYDKAKYNLLDYIYNNYEVDDDTILITNSDMGKGYTSRVFKELGKAL


KVKKHEHFWDIYHVKEKLSSYLRKYPIELTDFALDAVKKYNSDKLELVFDTVESLICDELEDQEFQKFKKKV


LNNFKYIKPAHLRNLSNRGIGIMESQHRKITYRMKRRGMYWSKWGISTMANMIILERANGLRELFFGSWRKV


YSEYKEGSFSAGRLFKKTDELDKFSKPLLKNGRKWSITGIKTK


 


Strain COH1


ORF DNA sequence (SEQ ID NO: 88):


ATGGAAGTTAAAAAATTCTCGGAAAAAGATTTTGTAAATGAAATAAATAAAATAAAACAGAAACAATTTTTA


AGTCAAATTGAACAGTATGAAAGCTATATCGCTCCTCAAATGAGAACGAAAGGCTATAAGAGGATCAATCAG


TCTGAACGTACGGTTGTTTTCTCTTTTGGAGAAATAACATTTAGTAGGAGTCGCTGGACAAATGGCTTTGAA


ACTAGAATACCAGTAGATGAGTGGTTAGGTCTTGAAAAATATAAGAGATATTCAATAGAATTCTTATATCAT


GTTGCAAAATTGGCTACAATGATGCCTTATCGTCAAGTTTGCAAAGTAATAGATAGCACTTTGCAAACAATC


ATAACAAAAGACTGTGTTTTAAAAGCAGTAAAATTTGTAGAAAAATTGTTAAAAGAAAAAGAACGCTATCGT


TTTTATTTGGAAGAGCCACCCGAACGTAAAAAAGTGAAAAAACTGTATGTTGAGGGTGATGGAGTCATGATT


AAAAGCACAGATTCTAGAGAGGAAAGAAGGTATTTAGATTTAACACATTTTGTTATTCATACAGGCTCAAAA


AAAGTTTCTACTAAAAGATATGAATTGCGGGACAAGCACGAAATATTACAGCTTAATTATGATAAAGCTAAA


TATAATCTTTTAGATTATATTTATAATAACTATGAAGTAGATGACGATACTATTTTAATCACTAACTCTGAT


ATGGGTAAAGGCTATACTAGTAGAGTTTTTAAGGAATTAGGAAAAGCACTTAAGGTAAAGAAACATGAGCAT


TTTTGGGATATCTATCATGTTAAAGAAAAGTTAAGTTCATACCTTAGAAAATATCCAATTGAATTAACCGAT


TTTGCTTTAGATGCGGTAAAAAAATATAATTCTGATAAGCTTGAATTAGTTTTTGATACTGTTGAATCACTG


ATTTGTGATGAACTTGAAGATCAAGAATTTCAGAAGTTTAAGAAAAAAGTATTAAATAATTTCAAATATATA


AAACCAGCTCATCTTAGAAATCTTTCAAATCGTGGTATTGGTATCATGGAATCACAACACAGAAAGATAACG


TATAGAATGAAGCGACGTGGCATGTATTGGTCAAAGTGGGGAATCTCCACAATGGCAAATATGATTATACTT


GAAAGAGCTAACGGTTTACGAGAATTATTTTTCGGTTCTTGGAGAAAGGTATACAGTGAGTATAAAGAAGGT


TCATTTAGTGCAGGGCGACTTTTTAAAAAGACAGATGAATTAGATAAATTTTCTAAGCCCCTTCTAAAAAAT


GGCAGAAAATGGAGTATAACAGGAATCAAAACAAAATAG


 


ORF amino acid sequence (SEQ ID NO: 82):


MEVKKFSEKDFVNEINKIKQKQFLSQIEQYESYIAPQMRTKGYKRINQSERTVVFSFGEITFSRSRWTNGFE


TRIPVDEWLGLEKYKRYSIEFLYHVAKLATMMPYRQVCKVIDSTLQTIITKDCVLKAVKFVEKLLKEKERYR


FYLEEPPERKKVKKLYVEGDGVMIKSTDSREERRYLDLTHFVIHTGSKKVSTKRYELRDKHEILQLNYDKAK


YNLLDYIYNNYEVDDDTILITNSDMGKGYTSRVFKELGKALKVKKHEHFWDIYHVKEKLSSYLRKYPIELTD


FALDAVKKYNSDKLELVFDTVESLICDELEDQEFQKFKKKVLNNFKYIKPAHLRNLSNRGIGIMESQHRKIT


YRMKRRGMYWSKWGISTMANMIILERANGLRELFFGSWRKVYSEYKEGSFSAGRLFKKTDELDKFSKPLLKN


GRKWSITGIKTK


 


Strain IC105


ORF DNA sequence (SEQ ID NO: 89):


TTTAGTGTAACCTATTCACAGTCTGAACGTACGGTTGTTTTCTCTTTTGGAGAAATAACATTTAGTAGGAGT


CGCTGGACAAATGGCTTTGAAACTAGAATACCAGTAGATGAGTGGTTAGGTCTTGAAAAATATAAGAGATAT


TCAATAGAATTCTTATATCATGTTGCAAAATTGGCTACAATGATGCCTTATCGTCAAGTTTGCAAAGTAATA


GATAGCACTTTGCAAACAATCATAACAAAAGACTGTGTTTTAAAAGCAGTAAAATTTGTAGAAAAATTGTTA


AAAGAAAAAGAACGCTATCGTTTTTATTTGGAAGAGCCACCCGAACGTAAAAAAGTGAAAAAACTGTATGTT


GAGGGTGATGGAGTCATGATTAAAAGCACAGATTCTAGAGAGGAAAGAAGGTATTTAGATTTAACACATTTT


GTTATTCATACAGGCTCAAAAAAAGTTTCTACTAAAAGATATGAATTGCAGGACAAGCACGAAATATTACAG


CTTAATTATGATAAAGCTAAATATAATCTTTTAGATTATATTTATAATAACTATGAAGTAGATGACGATACT


ATTTTAATCACTAACTCTGATATGGGTAAAGGCTATACTAGTAGAGTTTTTAAGGAATTAGGAAAAGCACTT


AAGGTAAAGAAACATGAGCATTTTTGGGATATCTATCATGTTAAAGAAAAGTTAAGTTCATACCTTAGAAAA


TATCCAATTGAATTAACCGATTTTGCTTTAGATGCGGTAAAAAAATATAATTCTGATAAGCTTGAATTAGTT


TTTGATACTGTTGAATCACTGATTTGTGATGAACTTGAAGATCAAGAATTTCAGAAGTTTAAGAAAAAAGTA


TTAAATAATTTCAAATATATAAAACCAGCTCATCTTAGAAATCTTTCAAATCGTGGTATTGGTATCATGGAA


TCACAACACAGAAAGATAACGTATAGAATGAAGCGACGTGGCATGTATTGGTCAAAGTGGGGAATCTCCACA


ATGGCAAATATGATTATACTTGAAAGAGCTAACGGTTTACGAGAATTATTTTTCGGTTCTTGGAGAAAGGTA


TACAGTGAGTATAAAGAAGGTTCATTTAGTGCAGGGCGACTTTTTAAAAAGACAGATGAATTAGATAAATTT


TCTAAGCCCCTTCTAAAAAATGGCAGAAAATGGAGTATAACAGGAATCAAAACAAAATAG


 


ORF amino acid sequence (SEQ ID NO: 83):


FSVTYSQSERTVVFSFGEITFSRSRWTNGFETRIPVDEWLGLEKYKRYSIEFLYHVAKLATMMPYRQVCKVI


DSTLQTIITKDCVLKAVKFVEKLLKEKERYRFYLEEPPERKKVKKLYVEGDGVMIKSTDSREERRYLDLTHF


VIHTGSKKVSTKRYELQDKHEILQLNYDKAKYNLLDYIYNNYEVDDDTILITNSDMGKGYTSRVFKELGKAL


KVKKHEHFWDIYHVKEKLSSYLRKYPIELTDFALDAVKKYNSDKLELVFDTVESLICDELEDQEFQKFKKKV


LNNFKYIKPAHLRNLSNRGIGIMESQHRKITYRMKRRGMYWSKWGISTMANMIILERANGLRELFFGSWRKV


YSEYKEGSFSAGRLFKKTDELDKFSKPLLKNGRKWSITGIKTK


 


Strain IC458


ORF DNA sequence (SEQ ID NO: 90):


TTTAGTGTAACCTATTCACAGTCTGAACGTACGGTTGTTTTCTCTTTTGGAGAAATAACATTTAGTAGGAGT


CGCTGGACAAATGGCTTTGAAACTAGAATACCAGTAGATGAGTGGTTAGGTCTTGAAAAATATAAGAGATAT


TCAATAGAATTCTTATATCATGTTGCAAAATTGGCTACAATGATGCCTTATCGTCAAGTTTGCAAAGTAATA


GATAGCACTTTGCAAACAATCATAACAAAAGACTGTGTTTTAAAAGCAGTAAAATTTGTAGAAAAATTGTTA


AAAGAAAAAGAACGCTATCGTTTTTATTTGGAAGAGCCACCCGAACGTAAAAAAGTGAAAAAACTGTATGTT


GAGGGTGATGGAGTCATGATTAAAAGCACAGATTCTAGAGAGGAAAGAAGGTATTTAGATTTAACACATTTT


GTTATTCATACAGGCTCAAAAAAAGTTTCTACTAAAAGATATGAATTGCAGGACAAGCACGAAATATTACAG


CTTAATTATGATAAAGCTAAATATAATCTTTTAGATTATATTTATAATAACTATGAAGTAGATGACGATACT


ATTTTAATCACTAACTCTGATATGGGTAAAGGCTATACTAGTAGAGTTTTTAAGGAATTAGGAAAAGCACTT


AAGGTAAAGAAACATGAGCATTTTTGGGATATCTATCATGTTAAAGAAAAGTTAAGTTCATACCTTAGAAAA


TATCCAATTGAATTAACCGATTTTGCTTTAGATGCGGTAAAAAAATATAATTCTGATAAGCTTGAATTAGTT


TTTGATACTGTTGAATCACTGATTTGTGATGAACTTGAAGATCAAGAATTTCAGAAGTTTAAGAAAAAAGTA


TTAAATAATTTCAAATATATAAAACCAGCTCATCTTAGAAATCTTTCAAATCGTGGTATTGGTATCATGGAA


TCACAACACAGAAAGATAACGTATAGAATGAAGCGACGTGGCATGTATTGGTCAAAGTGGGGAATCTCCACA


ATGGCAAATATGATTATACTTGAAAGAGCTAACGGTTTACGAGAATTATTTTTCGGTTCTTGGAGAAAGGTA


TACAGTGAGTATAAAGAAGGTTCATTTAGTGCAGGGCGACTTTTTAAAAAGACAGATGAATTAGATAAATTT


TCTAAGCCCCTTCTAAAAAATGGCAGAAAATGGAGTATAACAGGAATCAAAACAAAATAG


 


ORF amino acid sequence (SEQ ID NO: 84):


FSVTYSQSERTVVFSFGEITFSRSRWTNGFETRIPVDEWLGLEKYKRYSIEFLYHVAKLATMMPYRQVCKVI


DSTLQTIITKDCVLKAVKFVEKLLKEKERYRFYLEEPPERKKVKKLYVEGDGVMIKSTDSREERRYLDLTHF


VIHTGSKKVSTKRYELQDKHEILQLNYDKAKYNLLDYIYNNYEVDDDTILITNSDMGKGYTSRVFKELGKAL


KVKKHEHFWDIYHVKEKLSSYLRKYPIELTDFALDAVKKYNSDKLELVFDTVESLICDELEDQEFQKFKKKV


LNNFKYIKPAHLRNLSNRGIGIMESQHRKITYRMKRRGMYWSKWGISTMANMIILERANGLRELFFGSWRKV


YSEYKEGSFSAGRLFKKTDELDKFSKPLLKNGRKWSITGIKTK
















TABLE 10





Amino acid and encoding DNA sequences of gbs1477 proteins derived from


different strains of S. agalactiae







Strain 0176H4A


ORF DNA sequence (SEQ ID NO: 133):


ATGAAAAAAATCAACAAATTTTTTGTGGCGTTCTCAGCGTTGTTACTGATTTTAACGTCATTGCTCTCAGTT


GCACCAGCGTTTGCGGAAAAAGAAAAAACAACTGAGACTGTTACTTTGCATAAAATTTTACAAACTGATACA


AACCTTAAGAATAGTGCTTTCCCTGGTACAAAAGGGCTAGATGGAACTGAATATGACGGGAAAGCTATTGAT


AAATTGGATAGCTACTTTGGCAATGACTCAAAAGATATTGGTGGGGCTTACTTTATATTGGCAAATAGCAAG


GGTGAATATATCAAAGCTAATGATAAAAATAAATTAAAGCCTGAGTTTAGTGGGAACACTCCGAAAACGACC


CTCAATATTAGTGAAGCTGTAGGTGGTTTGACAGAAGAAAACGCAGGTATTAAGTTTGAAACCACTGGTTTA


AGAGGGGATTTCCAGATTATTGAATTGAAAGACAAGTCAACTTACAATAATGGTGGGGCCATCTTGGCTGAT


TCAAAAGCGGTTCCAGTGAAAATCACTCTTCCATTGATAAACAAGGATGGTGTTGTTAAAGATGCACACGTC


TATCCAAAGAACACTGAAACAAAACCGCAAATTGACAAGAACTTTGCTGATAAAAATCTTGATTATATTAAC


AACCAAAAAGACAAAGGTACTATATCAGCAACTGTTGGTGATGTTAAAAAATATACTGTTGGGACAAAAATC


CTTAAAGGATCTGACTATAAAAAATTAGTTTGGACCGATAGCATGACGAAAGGATTGACGTTTAACAACGAT


GTTACTGTAACATTGGATGGTGCAAATTTTGAACAATCAAATTACACCTTAGTAGCTGATGACCAAGGTTTC


CGTCTTGTCTTGAATGCAACAGGTCTTTCTAAAGTAGCAGAAGCTGCAAAAACAAAAGATGTTGAAATCAAA


ATCAACTATTCAGCTACAGTAAACGGTTCTACTGTCGTTGAAAAGTCAGAAAATAATGATGTCAAACTAGAT


TATGGTAACAACCCAACAACTGAAAACGAACCACAAACTGGTAATCCAGTTAACAAAGAAATCACAGTTCGA


AAGACTTGGGCAGTGGATGGTAATGAAGTGAATAAGGGAGATGAAAAAGTTGACGCTGTCTTCACGTTGCAA


GTTAAAGATAGTGACAAATGGGTGAATGTCGATTCAGCAACAGCAACAGCAGCAACTGACTTCAAATACACT


TTCAAAAACTTGGATAATGCCAAAACTTACCGTGTTGTAGAACGTGTTAGCGGCTACGCTCCAGCCTACGTT


TCATTTGTGGGTGGAGTTGTGACTATTAAGAATAACAAAAACTCAAATGACCCAACTCCAATCAATCCATCA


GAACCAAAAGTTGTGACTTATGGACGTAAATTTGTGAAAACAAATCAAGATGGCTCTGAACGTCTAGCAGGA


GCTACTTTCCTTGTTAAGAACTCACAAAGTCAATACTTGGCACGTAAATCAGGTGTTGCAACTAATGAAGCT


CACAAAGCAGTAACAGATGCTAAAGTACAACTGGATGAAGCTGTTAAAGCTTATAACAAATTGACTAAAGAA


CAACAAGAAAGTCAAGATGGTAAAGCAGCATTGAATCTTATTGATGAAAAACAAACAGCTTACAATGAAGCT


TTTGCTAAAGCAAACTACTCATATGAATGGGTTGTAGATAAAAACGCTGCAAACGTTGTTAAATTGATTTCT


AATACAGCTGGTAAATTTGAAATTACAGGTTTGAATGCAGGCGAGTATAGTTTGGAAGAGACTCAAGCACCA


ACAGGTTATGCTAAATTGTCAAGTGATGTATCATTTAAAGTAAATGATACATCGTATAGCGAAGGGGCTTCA


AATGATATTGCATACGATAAAGACTCCGGTAAAACAGATGCACAAAAAGTTGTCAACAAAAAAGTAACAATC


CCACAAACAGGTGGTATTGGTACAATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCGGTAGTT


ATCATGAAAAGACGTCAATCAGAGGAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 91):


MKKINKFFVAFSALLLILTSLLSVAPAFAEKEKTTETVTLHKILQTDTNLKNSAFPGTKGLDGTEYDGKAID


KLDSYFGNDSKDIGGAYFILANSKGEYIKANDKNKLKPEFSGNTPKTTLNISEAVGGLTEENAGIKFETTGL


RGDFQIIELKDKSTYNNGGAILADSKAVPVKITLPLINKDGVVKDAHVYPKNTETKPQIDKNFADKNLDYIN


NQKDKGTISATVGDVKKYTVGTKILKGSDYKKLVWTDSMTKGLTFNNDVTVTLDGANFEQSNYTLVADDQGF


RLVLNATGLSKVAEAAKTKDVEIKINYSATVNGSTVVEKSENNDVKLDYGNNPTTENEPQTGNPVNKEITVR


KTWAVDGNEVNKGDEKVDAVFTLQVKDSDKWVNVDSATATAATDFKYTFKNLDNAKTYRVVERVSGYAPAYV


SFVGGVVTIKNNKNSNDPTPINPSEPKVVTYGRKFVKTNQDGSERLAGATFLVKNSQSQYLARKSGVATNEA


HKAVTDAKVQLDEAVKAYNKLTKEQQESQDGKAALNLIDEKQTAYNEAFAKANYSYEWVVDKNAANVVKLIS


NTAGKFEITGLNAGEYSLEETQAPTGYAKLSSDVSFKVNDTSYSEGASNDIAYDKDSGKTDAQKVVNKKVTI


PQTGGIGTILFTIIGLSIMLGAVVIMKRRQSEEA


 


Strain 12351


ORF DNA sequence (SEQ ID NO: 134):


ATGAAAAAAATCAACAAATATTTTGCAGTCTTCTCGGCATTGCTACTGACCGTAACATCATTGTTCTCAGTT


GCACCAGTGTTTGCGGAAGAAGCAAAAACTACTGACACAGTGACCTTGCACAAGATTGTCATGCCTCGAACT


GCATTTGACGGTTTTACTGCTGGTACAAAGGGTAAGGATAATACTGACTACGTTGGTAAACAAATCGAAGAC


CTTAAAACTTACTTTGGCTCAGGCGAAGCGAAAGAAATCGCAGGTGCTTACTTTGCTTTCAAAAATGAAGCT


GGTACTAAATACATCACTGAAAATGGTGAAGAAGTTGATACTTTGGATACAACAGATGCCAAAGGTGGTGCT


GTTCTTAAAGGTTTAACAACAGACAATGGTTTCAAATTTAACACTTCTAAATTAACAGGAACTTACCAAATC


GTTGAATTGAAAGAAAAATCTACATACAACAACGATGGTTCTATCTTGGCTGATTCAAAAGCAGTTCCAGTT


AAAATCACTCTTCCATTGGTAAACGACAATGGTGTTGTTAAAGACGCTCACGTTTATCCAAAGAACACTGAA


ACAAAACCACAAGTAGATAAGAACTTCGCAGATAAAGAACTTGATTATGCGAACAACAAAAAAGACAAAGGG


ACTGTCTCAGCATCTGTTGGTGATGTTAAAAAATATCATGTTGGAACAAAAATCCTTAAAGGTTCAGACTAT


AAGAAATTAATCTGGACCGATAGCATGACCAAAGGTTTGACTTTCAACAACGATATTGCTGTAACATTGGAT


GGTGCAACTCTTGATGCTACAAATTACAAACTTGTAGCAGATGACCAAGGTTTCCGCCTTGTCTTGACTGAC


AAAGGTCTTGAAGCAGTGGCAAAAGCCGCAAAAACAAAAGATGTTGAAATCAAGATCACTTACTCAGCTACT


TTGAACGGTTCTGCTGTCGTTGAAGTTCTAGAAACCAATGATGTTAAATTGGACTACGGCAACAACCCAACA


ATTGAAAATGAACCAAAAGAAGGTATTCCAGTTGATAAGAAAATCACTGTTAACAAAACATGGGCAGTAGAT


GGCAATGAAGTGAATAAAGCAGATGAAACAGTTGATGCTGTCTTCACCTTGCAAGTTAAAGATGGTGACAAA


TGGGTGAATGTTGATTCAGCTAAAGCAACAGCTGCAACTAGCTTCAAACACACTTTTGAAAACTTGGATAAT


GCTAAAACTTACCGCGTTATCGAACGTGTTAGCGGCTACGCTCCAGAATACGTCTCATTTGTAAATGGCGTT


GTAACCATCAAGAACAACAAAGACTCAAATGAGCCAACTCCAATCAACCCATCAGAACCAAAAGTGGTGACT


TATGGACGTAAATTTGTGAAAACAAATAAAGATGGAAAAGAACGCTTGGCAGGAGCTACCTTCCTTGTTAAG


AAAGATGGCAAGTACTTGGCACGTAAATCAGGTGTTGCAACAGATGCAGAAAAAGCTGCTGTAGATTCAACT


AAATCAGCATTGGATGCTGCTGTTAAAGCTTACAATGATTTGACTAAAGAAAAACAAGAAGGTCAAGATGGT


AAATCAGCATTGGCTACCGTTAGTGAAAAACAAAAAGCTTACATTGATGCCTTTGTTAAAGCTAACTACTCA


TACGAATGGGTTGAAGATAAAAATGCTAAGAATGTTGTTAAATTGATTTCTAACGATAAAGGTCAATTTGAA


ATTACTGGCTTGACTGAAGGTCAATACTCATTGGAAGAAACACAAGCACCAACTGGTTATGCTAAATTATCA


GGTGATGTTTCGTTTAATGTTAATGCTACTTCATACAGTAAAGGTTCTGCTCAAGATATTGAGTATACCCAA


GGTTCTAAAACTAAAGATGCACAACAAGTTATCAATAAGAAGGTTACTATTCCACAAACAGGTGGTATTGGT


ACAATTTTTTTCACAATTATTGGATTAAGTATTATGCTTGGAGCGGTAGTTATCATGAAAAGACGTCAATCA


GAGGAAGTTTAA


 


ORF amino acid sequence (SEQ ID NO: 92):


MKKINKYFAVFSALLLTVTSLFSVAPVFAEEAKTTDTVTLHKIVMPRTAFDGFTAGTKGKDNTDYVGKQIED


LKTYFGSGEAKEIAGAYFAFKNEAGTKYITENGEEVDTLDTTDAKGGAVLKGLTTDNGFKFNTSKLTGTYQI


VELKEKSTYNNDGSILADSKAVPVKITLPLVNDNGVVKDAHVYPKNTETKPQVDKNFADKELDYANNKKDKG


TVSASVGDVKKYHVGTKILKGSDYKKLIWTDSMTKGLTFNNDIAVTLDGATLDATNYKLVADDQGFRLVLTD


KGLEAVAKAAKTKDVEIKITYSATLNGSAVVEVLETNDVKLDYGNNPTIENEPKEGIPVDKKITVNKTWAVD


GNEVNKADETVDAVFTLQVKDGDKWVNVDSAKATAATSFKHTFENLDNAKTYRVIERVSGYAPEYVSFVNGV


VTIKNNKDSNEPTPINPSEPKVVTYGRKFVKTNKDGKERLAGATFLVKKDGKYLARKSGVATDAEKAAVDST


KSALDAAVKAYNDLTKEKQEGQDGKSALATVSEKQKAYIDAFVKANYSYEWVEDKNAKNVVKLISNDKGQFE


ITGLTEGQYSLEETQAPTGYAKLSGDVSFNVNATSYSKGSAQDIEYTQGSKTKDAQQVINKKVTIPQTGGIG


TIFFTIIGLSIMLGAVVIMKRRQSEEV


 


Strain 12401


ORF DNA sequence (SEQ ID NO: 135):


ATGAAAAGAATCAACAAATATTTTGCAATGTTCTCGGCATTGTTATTAATTTTAACATCGTTGTTATCGGTA


GCTCCGGTATTTGCTGCTGAGATGGGAAATATCACTAAAACAGTAACCTTACACAAAATTGTTCAAACATCC


GATAATTTGGCTAAGCCAAATTTCCCAGGAATAAATGGATTGAATGGAACGAAGTATATGGGTCAAAAACTT


ACTGACATTTCAGGATATTTTGGGCAAGGTTCTAAAGAAATCGCCGGTGCTTTCTTTGCGGTTATGAATGAA


AGTCAGACAAAATATATCACAGAAAGTGGTACTGAAGTAGAAAGTATCGATGCAGCAGGTGTCCTTAAAGGT


TTGACAACTGAAAACGGCATTACATTTAATACTGCAAACTTAAAAGGAACATACCAAATCGTTGAGTTGCTT


GACAAATCTAATTATAAAAATGGTGACAAAGTTCTTGCTGACTCAAAAGCTGTCCCAGTGAAAATCACTCTT


CCTTTGTATAACGAAGAAGGAATTGTCGTGGACGCTGAAGTGTATCCAAAGAATACAGAAGAAGCACCACAA


ATCGACAAAAACTTTGCTAAAGCAAATAAATTGTTGAATGACAGTGATAATTCAGCTATTGCAGGTGGGGCA


GACTACGACAAATATCAGGCAGAAAAAGCAAAAGCTACTGCTGAAATCGGTCAAGAAATCCCTTACGAAGTT


AAAACAAAAATCCAAAAAGGGTCTAAATACAAAAACCTTGCTTGGGTCGATACCATGTCAAATGGTTTGACA


ATGGGTAACACTGTTAACTTAGAAGCATCGTCAGGCTCTTTTGTAGAAGGTACAGATTACAATGTTGAACGT


GATGACCGTGGTTTCACTTTGAAATTCACAGATACAGGTTTGACTAAGCTACAAAAAGAAGCGGAAACACAA


GCTGTTGAATTCACATTGACATATAGCGCAACAGTTAACGGTGCGGCTATTGATGACAAGCCAGAAAGCAAT


GATATCAAACTTCAATACGGTAACAAACCAGGTAAAAAAGTAAAAGAAATCCCAGTAACACCGTCAAATGGC


GAAATCACTGTTAGCAAAACTTGGGACAAAGGTTCAGATTTAGAGAATGCGAATGTTGTTTATACCCTTAAA


GATGGTGGAACAGCTGTTGCCTCAGTTTCATTGACAAAAACAACACCAAATGGCGAAATCAACTTAGGTAAT


GGTATTAAATTTACAGTTACTGGAGCGTTTGCTGGTAAATTCAGTGGTCTGACTGATAGTAAAACATACATG


ATCTCAGAACGTATCGCTGGTTATGGTAATACAATCACTACTGGTGCTGGTAGTGCAGCTATCACCAATACT


CCAGATTCAGACAACCCAACACCACTTAATCCAACTGAACCAAAAGTTGTGACACACGGTAAAAAATTCGTC


AAAACAAGTTCGACTGAAACAGAACGCTTGCAAGGTGCACAGTTCGTTGTTAAAGATTCAGCTGGTAAATAC


CTTGCATTGAAATCATCTGCGACAATATCAGCTCAAACAACAGCTTACACAAATGCTAAAACTGCTCTTGAC


GCTAAAATCGCAGCTTACAACAAACTTTCAGCAGACGATCAAAAAGGTACTAAAGGTGAAACAGCTAAAGCA


GAAATCAAAACTGCTCAAGACGCTTACAATGCAGCCTTCATCGTAGCTCGTACAGCTTACGAGTGGGTAACT


AATAAAGAAGATGCTAACGTTGTTAAAGTGACTTCAAACGCTGACGGTCAATTTGAAGTTAGCGGTCTTGCA


ACTGGTGATTATAAACTTGAAGAAACACAAGCTCCAGCTGGTTACGCTAAATTAGCAGGTGATGTTGATTTC


AAAGTTGGAAACAGCTCAAAAGCAGACGACTCAGGTAACATTGATTACACTGCTAGCAGCAATAAAAAAGAC


GCTCAACGCATAGAAAACAAAAAAGTGACTATTCCACAAACAGGTGGTATTGGTACAATTCTTTTCACAATT


ATTGGTTTAAGCATTATGCTTGGAGCGGTAATTATCATGAAAAGACGTCAATCAGAGGAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 93):


MKRINKYFAMFSALLLILTSLLSVAPVFAAEMGNITKTVTLHKIVQTSDNLAKPNFPGINGLNGTKYMGQKL


TDISGYFGQGSKEIAGAFFAVMNESQTKYITESGTEVESIDAAGVLKGLTTENGITFNTANLKGTYQIVELL


DKSNYKNGDKVLADSKAVPVKITLPLYNEEGIVVDAEVYPKNTEEAPQIDKNFAKANKLLNDSDNSAIAGGA


DYDKYQAEKAKATAEIGQEIPYEVKTKIQKGSKYKNLAWVDTMSNGLTMGNTVNLEASSGSFVEGTDYNVER


DDRGFTLKFTDTGLTKLQKEAETQAVEFTLTYSATVNGAAIDDKPESNDIKLQYGNKPGKKVKEIPVTPSNG


EITVSKTWDKGSDLENANVVYTLKDGGTAVASVSLTKTTPNGEINLGNGIKFTVTGAFAGKFSGLTDSKTYM


ISERIAGYGNTITTGAGSAAITNTPDSDNPTPLNPTEPKVVTHGKKFVKTSSTETERLQGAQFVVKDSAGKY


LALKSSATISAQTTAYTNAKTALDAKIAAYNKLSADDQKGTKGETAKAEIKTAQDAYNAAFIVARTAYEWVT


NKEDANVVKVTSNADGQFEVSGLATGDYKLEETQAPAGYAKLAGDVDFKVGNSSKADDSGNIDYTASSNKKD


AQRIENKKVTIPQTGGIGTILFTIIGLSIMLGAVIIMKRRQSEEA


 


Strain 126H4A


ORF DNA sequence (SEQ ID NO: 136):


ATGAAAAAAATCAACAAATATTTTGCAGTCTTCTCGGCCTTGCTACTGACCGTAACATCATTGCTCTCAGTT


GCACCAGCGTTTGCGGACGAAGCAACAACTAATACAGTGACTTTGCACAAGATCTTGCAAACTGAATCAAAT


CTTAATAAAAGTAACTTCCCAGGAACTACAGGCCTTAACGGAGATGACTATAAAGGTGAATCTATTTCTGAC


CTTGCTGAATACTTTGGATCAGGTTCTAAAGAAATTGACGGTGCTTTCTTTGCTTTGGCTTTAGAAGAGGAA


AAAGATGGTGTCGTACAATATGTTAAGGCAAAAGCAAATGACAAATTAACACCAGACTTAATTACTAAAGGT


ACACCTGCAACAACAACAAAAGTTGAAGAAGCTGTAGGTGGTTTGACAACTGGTACGGGTATTGTTTTCAAT


ACAGCTGGTTTGAAAGGTAATTTCAAAATTATTGAATTGAAAGACAAATCAACTTACAACAATAATGGTTCC


CTCTTAGCAGCTTCAAAAGCAGTTCCGGTGAAAATCACTCTTCCATTGGTAAGCAAAGATGGTGTTGTTAAA


GATGCACACGTTTATCCAAAGAACACTGAAACAAAACCAGAAGTAGACAAGAACTTCGCTAAAACAAACGAT


TTGACAGCTCTCAAAGACGCTACTCTTCTTAAGGCTGGTGCAGACTACAAAAACTATTCAGCGACTAAAGCT


ACTGTAACAGCTGAAATCGGTAAAGTTATCCCTTACGAAGTTAAAACAAAAGTTCTTAAAGGTTCTAAATAC


GAAAAACTGGTTTGGACCGATACCATGTCAAATGGTTTGACAATGGGTGATGATGTTAACCTTGCAGTTTCA


GGGACTACAACAACTTTCATTAAAGATATAGATTACACTCTTAGCATTGATGACCGTGGTTTCACATTGAAA


TTCAAAGCTACTGGATTGGACAAATTGGAAGAAGCAGCTAAAGCATCTGATGTTGAATTTACATTGACTTAT


AAAGCTACTGTTAATGGCCAAGCAATTATTGACAACCCAGAAGTCAATGACATCAAATTGGACTATGGTAAT


AAACCTGGTACAGATTTATCAGAACAACCTGTGACACCTGAAGATGGTGAAGTTAAAGTCACTAAAACATGG


GCAGCAGGTGCTAATAAAGCAGACGCTAAAGTTGTCTACACACTTAAAAATGCTACTAAACAAGTCGTAGCT


TCTGTCGCATTGACCGCAGCTGATACAAAAGGTACGATTAATCTTGGTAAAGGCATGACCTTTGAAATCACA


GGAGCTTTCTCAGGTACATTCAAAGGCCTTCAAAATAAAGCTTACACTGTTTCTGAACGTGTTGCAGGTTAT


ACTAATGCTATTAATGTTACTGGTAATGCTGTTGCTATCACCAATACACCAGACAGTGACAATCCAACGCCA


CTTAACCCAACTCAACCAAAAGTTGAAACACATGGTAAGAAATTTGTCAAAGTTGGCGATGCAGATGCCCGC


TTAGCTGGTGCACAATTCGTTGTGAAAAATTCAGCTGGTAAATTCCTTGCTCTTAAAGAAGATGCAGCTGTA


TCAGGAGCTCAAACTGAATTGGCAACTGCTAAAACAGACTTGGATAATGCCATCAAAGCTTACAACGGTTTG


ACAAAAGCGCAGCAAGAAGGTGCTGATGGTACATCAGCAAAAGAACTTATCAACACTAAACAGTCAGCTTAC


GACGCAGCCTTCATCAAAGCACGTACAGCTTATATATGGGTAGATGAAAAAACTAAAGCTATTACCTTCACT


TCAAATAATCAAGGTCAATTTGAAGTTACTGGTCTTGAAGTAGGTTCTTACAAACTTGAAGAAACTCTTGCA


CCAGCAGGTTATGCTAAATTGTCAGGCGACATTGAGTTTACAGTTGGACACGATTCTTACACAAGTGGTGAC


ATCAAGTACAAGACAGATGATGCTAGCAACAATGCACAAAAAGTTTTCAATAAAAAAGTAACCATCCCACAA


ACAGGTGGTATTGGTACAATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCGGTAGTTATCATG


AAAAGACGTCAATCAGAGGAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 94):


MKKINKYFAVFSALLLTVTSLLSVAPAFADEATTNTVTLHKILQTESNLNKSNFPGTTGLNGDDYKGESISD


LAEYFGSGSKEIDGAFFALALEEEKDGVVQYVKAKANDKLTPDLITKGTPATTTKVEEAVGGLTTGTGIVFN


TAGLKGNFKIIELKDKSTYNNNGSLLAASKAVPVKITLPLVSKDGVVKDAHVYPKNTETKPEVDKNFAKTND


LTALKDATLLKAGADYKNYSATKATVTAEIGKVIPYEVKTKVLKGSKYEKLVWTDTMSNGLTMGDDVNLAVS


GTTTTFIKDIDYTLSIDDRGFTLKFKATGLDKLEEAAKASDVEFTLTYKATVNGQAIIDNPEVNDIKLDYGN


KPGTDLSEQPVTPEDGEVKVTKTWAAGANKADAKVVYTLKNATKQVVASVALTAADTKGTINLGKGMTFEIT


GAFSGTFKGLQNKAYTVSERVAGYTNAINVTGNAVAITNTPDSDNPTPLNPTQPKVETHGKKFVKVGDADAR


LAGAQFVVKNSAGKFLALKEDAAVSGAQTELATAKTDLDNAIKAYNGLTKAQQEGADGTSAKELINTKQSAY


DAAFIKARTAYIWVDEKTKAITFTSNNQGQFEVTGLEVGSYKLEETLAPAGYAKLSGDIEFTVGHDSYTSGD


IKYKTDDASNNAQKVFNKKVTIPQTGGIGTILFTIIGLSIMLGAVVIMKRRQSEEA


 


Strain 49447


ORF DNA sequence (SEQ ID NO: 137):


ATGAAAAAAATCAACAAATTTTTTGTGGCGTTCTCAGCGTTGTTACTGATTTTAACGTCATTGCTCTCAGTT


GCACCAGCGTTTGCGGAAAAAGAAAAAACAACTGAGACTGTTACTTTGCATAAAATTTTACAAACTGATACA


AACCTTAAGAATAGTGCTTTCCCTGGTACAAAAGGGCTAGATGGAACTGAATATGACGGGAAAGCTATTGAT


AAATTGGATAGCTACTTTGGCAATGACTCAAAAGATATTGGTGGGGCTTACTTTATATTGGCAAATAGCAAG


GGTGAATATATCAAAGCTAATGATAAAAATAAATTAAAGCCTGAGTTTAGTGGGAACACTCCGAAAACGACC


CTCAATATTAGTGAAGCTGTAGGTGGTTTGACAGAAGAAAACGCAGGTATTAAGTTTGAAACCACTGGTTTA


AGAGGGGATTTCCAGATTATTGAATTGAAAGACAAGTCAACTTACAATAATGGTGGGGCCATCTTGGCTGAT


TCAAAAGCGGTTCCAGTGAAAATCACTCTTCCATTGATAAACAAGGATGGTGTTGTTAAAGATGCACACGTC


TATCCAAAGAACACTGAAACAAAACCGCAAATTGACAAGAACTTTGCTGATAAAAATCTTGATTATATTAAC


AACCAAAAAGACAAAGGTACTATATCAGCAACTGTTGGTGATGTTAAAAAATATACTGTTGGGACAAAAATC


CTTAAAGGATCTGACTATAAAAAATTAGTTTGGACCGATAGCATGACGAAAGGATTGACGTTTAACAACGAT


GTTACTGTAACATTGGATGGTGCAAATTTTGAACAATCAAATTACACCTTAGTAGCTGATGACCAAGGTTTC


CGTCTTGTCTTGAATGCAACAGGTCTTTCTAAAGTAGCAGAAGCTGCAAAAACAAAAGATGTTGAAATCAAA


ATCAACTATTCAGCTACAGTAAACGGTTCTACTGTCGTTGAAAAGTCAGAAAATAATGATGTCAAACTAGAT


TATGGTAACAACCCAACAACTGAAAACGAACCACAAACTGGTAATCCAGTTAACAAAGAAATCACAGTTCGA


AAGACTTGGGCAGTGGATGGTAATGAAGTGAATAAGGGAGATGAAAAAGTTGACGCTGTCTTCACGTTGCAA


GTTAAAGATAGTGACAAATGGGTGAATGTCGATTCAGCAACAGCAACAGCAGCAACTGACTTCAAATACACT


TTCAAAAACTTGGATAATGCCAAAACTTACCGTGTTGTAGAACGTGTTAGCGGCTACGCTCCAGCCTACGTT


TCATTTGTGGGTGGAGTTGTGACTATTAAGAATAACAAAAACTCAAATGACCCAACTCCAATCAATCCATCA


GAACCAAAAGTTGTGACTTATGGACGTAAATTTGTGAAAACAAATCAAGATGGCTCTGAACGTCTAGCAGGA


GCTACTTTCCTTGTTAAGAACTCACAAAGTCAATACTTGGCACGTAAATCAGGTGTTGCAACTAATGAAGCT


CACAAAGCAGTAACAGATGCTAAAGTACAACTGGATGAAGCTGTTAAAGCTTATAACAAATTGACTAAAGAA


CAACAAGAAAGTCAAGATGGTAAAGCAGCATTGAATCTTATTGATGAAAAACAAACAGCTTACAATGAAGCT


TTTGCTAAAGCAAACTACTCATATGAATGGGTTGTAGATAAAAACGCTGCAAACGTTGTTAAATTGATTTCT


AATACAGCTGGTAAATTTGAAATTACAGGTTTGAATGCAGGCGAGTATAGTTTGGAAGAGACTCAAGCACCA


ACAGGTTATGCTAAATTGTCAAGTGATGTATCATTTAAAGTAAATGATACATCGTATAGCGAAGGGGCTTCA


AATGATATTGCATACGATAAAGACTCCGGTAAAACAGATGCACAAAAAGTTGTCAACAAAAAAGTAACAATC


CCACAAACAGGTGGTATTGGTACAATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCGGTAGTT


ATCATGAAAAGACGTCAATCAGAGGAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 95):


MKKINKFFVAFSALLLILTSLLSVAPAFAEKEKTTETVTLHKILQTDTNLKNSAFPGTKGLDGTEYDGKAID


KLDSYFGNDSKDIGGAYFILANSKGEYIKANDKNKLKPEFSGNTPKTTLNISEAVGGLTEENAGIKFETTGL


RGDFQIIELKDKSTYNNGGAILADSKAVPVKITLPLINKDGVVKDAHVYPKNTETKPQIDKNFADKNLDYIN


NQKDKGTISATVGDVKKYTVGTKILKGSDYKKLVWTDSMTKGLTFNNDVTVTLDGANFEQSNYTLVADDQGF


RLVLNATGLSKVAEAAKTKDVEIKINYSATVNGSTVVEKSENNDVKLDYGNNPTTENEPQTGNPVNKEITVR


KTWAVDGNEVNKGDEKVDAVFTLQVKDSDKWVNVDSATATAATDFKYTFKNLDNAKTYRVVERVSGYAPAYV


SFVGGVVTIKNNKNSNDPTPINPSEPKVVTYGRKFVKTNQDGSERLAGATFLVKNSQSQYLARKSGVATNEA


HKAVTDAKVQLDEAVKAYNKLTKEQQESQDGKAALNLIDEKQTAYNEAFAKANYSYEWVVDKNAANVVKLIS


NTAGKFEITGLNAGEYSLEETQAPTGYAKLSSDVSFKVNDTSYSEGASNDIAYDKDSGKTDAQKVVNKKVTI


PQTGGIGTILFTIIGLSIMLGAVVIMKRRQSEEA


 


Strain 5095S2


ORF DNA sequence (SEQ ID NO: 138):


ATGAAAAGAATCAACAAATATTTTGCAATGTTCTCGGCATTGTTATTAATTTTAACATCGTTGTTATCGGTA


GCTCCGGTATTTGCTGCTGAGATGGGAAATATCACTAAAACAGTAACCTTACACAAAATTGTTCAAACATCC


GATAATTTGGCTAAGCCAAATTTCCCAGGAATAAATGGATTGAATGGAACGAAGTATATGGGTCAAAAACTT


ACTGACATTTCAGGATATTTTGGGCAAGGTTCTAAAGAAATCGCCGGTGCTTTCTTTGCGGTTATGAATGAA


AGTCAGACAAAATATATCACAGAAAGTGGTACTGAAGTAGAAAGTATCGATGCAGCAGGTGTCCTTAAAGGT


TTGACAACTGAAAACGGCATTACATTTAATACTGCAAACTTAAAAGGAACATACCAAATCGTTGAGTTGCTT


GACAAATCTAATTATAAAAATGGTGACAAAGTTCTTGCTGACTCAAAAGCTGTCCCAGTGAAAATCACTCTT


CCTTTGTATAACGAAGAAGGAATTATCGTGGACGCTGAAGTGTATCCAAAGAATACAGAAGAAGCACCACAA


ATCGACAAAAACTTTGCTAAAGCAAATAAATTGTTGAATGACAGTGATAATTCAGCTATTGCAGGTGGGGCA


GACTACGACAAATATCAGGCAGAAAAAGCAAAAGCTACTGCTGAAATCGGTCAAGAAATCCCTTACGAAGTT


AAAACAAAAATCCAAAAAGGGTCTAAATACAAAAACCTTGCTTGGGTCGATACCATGTCAAATGGTTTGACA


ATGGGTAACACTGTTAACTTAGAAGCATCGTCAGGCTCTTTTGTAGAAGGTACAGATTACAATGTTGAACGT


GATGACCGTGGTTTCACTTTGAAATTCACAGATACAGGTTTGACTAAGCTACAAAAAGAAGCGGAAACACAC


GCTGTTGAATTCACATTGACATATAGCGCAACAGTTAACGGTGCGGCTATTGATGACAAGCCAGAAAGCAAT


GATATCAAACTTCAATACGGTAACAAACCAGGTAAAAAAGTAAAAGAAATCCCAGTAACACCGTCAAATGGC


GAAATCACTGTTAGCAAAACTTGGGACAAAGGTTCAGATTTAGAGAATGCGAATGTTGTTTATACCCTTAAA


GATGGTGGAACAGCTGTTGCCTCAGTTTCATTGACAAAAACAACACCAAATGGCGAAATCAACTTAGGTAAT


GGTATTAAATTTACAGTTACTGGAGCGTTTGCTGGTAAATTCAGTGGTCTGACTGATAGTAAAACATACATG


ATCTCAGAACGTATCGCTGGTTATGGTAATACAATCACTACTGGTGCTGGTAGTGCAGCTATCACCAATACT


CCAGATTCAGACAACCCAACACCACTTAATCCAACTGAACCAAAAGTTGTGACACACGGTAAAAAATTCGTC


AAAACAAGTTCGACTGAAACAGAACGCTTGCAAGGTGCACAGTTCGTTGTTAAAGATTCAGCTGGTAAATAC


CTTGCATTGAAATCATCTGCGACAATATCAGCTCAAACAACAGCTTACACAAATGCTAAAACTGCTCTTGAC


GCTAAAATCGCAGCTTACAACAAACTTTCAGCAGACGATCAAAAAGGTACTAAAGGTGAAACAGCTAAAGCA


GAAATCAAAACTGCTCAAGACGCTTACAATGCAGCCTTCATCGTAGCTCGTACAGCTTACGAGTGGGTAACT


AATAAAGAAGATGCTAACGTTGTTAAAGTGACTTCAAACGCTGACGGTCAATTTGAAGTTAGCGGTCTTGCA


ACTGGTGATTATAAACTTGAAGAAACACAAGCTCCAGCTGGTTACGCTAAATTAGCAGGTGATGTTGATTTC


AAAGTTGGAAACAGCTCAAAAGCAGACGACTCAGGTAACATTGATTACACTGCTAGCAGCAATAAAAAAGAC


GCTCAACGCATAGAAAACAAAAAAGTGACTATTCCACAAACAGGTGGTATTGGTACAATTCTTTTCACAATT


ATTGGTTTAAGCATTATGCTTGGAGCGGTAATTATCATGAAAAGACGTCAATCAGAGGAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 96):


MKRINKYFAMFSALLLILTSLLSVAPVFAAEMGNITKTVTLHKIVQTSDNLAKPNFPGINGLNGTKYMGQKL


TDISGYFGQGSKEIAGAFFAVMNESQTKYITESGTEVESIDAAGVLKGLTTENGITFNTANLKGTYQIVELL


DKSNYKNGDKVLADSKAVPVKITLPLYNEEGIIVDAEVYPKNTEEAPQIDKNFAKANKLLNDSDNSAIAGGA


DYDKYQAEKAKATAEIGQEIPYEVKTKIQKGSKYKNLAWVDTMSNGLTMGNTVNLEASSGSFVEGTDYNVER


DDRGFTLKFTDTGLTKLQKEAETHAVEFTLTYSATVNGAAIDDKPESNDIKLQYGNKPGKKVKEIPVTPSNG


EITVSKTWDKGSDLENANVVYTLKDGGTAVASVSLTKTTPNGEINLGNGIKFTVTGAFAGKFSGLTDSKTYM


ISERIAGYGNTITTGAGSAAITNTPDSDNPTPLNPTEPKVVTHGKKFVKTSSTETERLQGAQFVVKDSAGKY


LALKSSATISAQTTAYTNAKTALDAKIAAYNKLSADDQKGTKGETAKAEIKTAQDAYNAAFIVARTAYEWVT


NKEDANVVKVTSNADGQFEVSGLATGDYKLEETQAPAGYAKLAGDVDFKVGNSSKADDSGNIDYTASSNKKD


AQRIENKKVTIPQTGGIGTILFTIIGLSIMLGAVIIMKRRQSEEA


 


Strain 6313


ORF DNA sequence (SEQ ID NO: 139):


ATGAAAAAAATCAACAAATGTCTTACAGTGTTCTCGACACTGCTATTGATCTTAACGTCACTATTCTCAGTT


GCACCAGCGTTTGCGGACGACGTAACAACTGATACTGTGACCTTGCACAAGATTGTCATGCCACAAGCTGCA


TTTGATAACTTTACTGAAGGTACAAAAGGTAAGAATGATAGCGATTATGTTGGTAAACAAATTAATGACCTT


AAATCTTATTTTGGCTCAACCGATGCTAAAGAAATTAAGGGTGCTTTCTTTGTTTTCAAAAATGAAACTGGT


ACAAAATTCATTACTGAAAATGGTAAGGAAGTCGATACTTTGGAAGCTAAAGATGCTGAAGGTGGTGCTGTT


CTTTCAGGGTTAACAAAAGACACTGGTTTTGCTTTTAACACTGCTAAGTTAAAAGGAACTTACCAAATCGTT


GAATTGAAAGAAAAATCAAACTACGATAACAACGGTTCTATCTTGGCTGATTCAAAAGCAGTTCCAGTTAAA


ATCACTCTGCCATTGGTAAACAACCAAGGTGTTGTTAAAGATGCTCACATTTATCCAAAGAATACTGAAACA


AAACCACAAGTAGATAAGAACTTTGCAGATAAAGATCTTGATTATACTGACAACCGAAAAGACAAAGGTGTT


GTCTCAGCGACAGTTGGTGACAAAAAAGAATACATAGTTGGAACAAAAATTCTTAAAGGCTCAGACTATAAG


AAACTGGTTTGGACTGATAGCATGACTAAAGGTTTGACGTTCAACAACAACGTTAAAGTAACATTGGATGGT


AAAGATTTTCCTGTTTTAAACTACAAACTCGTAACAGATGACCAAGGTTTCCGTCTTGCCTTGAATGCAACA


GGTCTTGCAGCAGTAGCAGCTGCTGCAAAAGACAAAGATGTTGAAATCAAGATCACTTACTCAGCTACGGTG


AACGGCTCCACTACTGTTGAAGTTCCAGAAACCAATGATGTTAAATTGGACTATGGTAATAACCCAACGGAA


GAAAGTGAACCACAAGAAGGTACTCCAGCTAACCAAGAAATTAAAGTCATTAAAGACTGGGCAGTAGATGGT


ACAATTACTGATGTTAATGTTGCAGTTAAAGCTATCTTTACCTTGCAAGAAAAACAAACGGATGGTACATGG


GTGAACGTTGCTTCACACGAAGCAACAAAACCATCACGCTTTGAACATACTTTCACAGGTTTGGATAATACT


AAAACTTACCGCGTTGTCGAACGTGTTAGCGGCTACACTCCAGAATATGTATCATTTAAAAATGGTGTTGTG


ACTATCAAGAACAACAAAAACTCAAATGATCCAACTCCAATCAACCCATCAGAACCAAAAGTGGTGACTTAT


GGACGTAAATTTGTGAAAACAAATCAAGCTAACACTGAACGCTTGGCAGGAGCTACCTTCCTTGTTAAGAAA


GAAGGAAAATACTTGGCACGTAAAGCAGGTGCAGCAACTGCTGAAGCAAAGGCAGCTGTAAAAACTGCTAAA


CTAGCATTGGATGAAGCTGTTAAAGCTTATAACGACTTGACTAAAGAAAAACAAGAAGGCCAAGAAGGTAAA


ACAGCATTGGCTACTGTTGATCAAAAACAAAAAGCTTACAATGACGCTTTTGTTAAAGCTAACTACTCATAT


GAATGGGTTGCAGATAAAAAGGCTGATAATGTTGTTAAATTGATCTCTAACGCCGGTGGTCAATTTGAAATT


ACTGGTTTGGATAAAGGCACTTATAGCTTGGAAGAAACTCAAGCACCAGCAGGTTATGCGACATTGTCAGGT


GATGTAAACTTTGAAGTAACTGCCACATCATATAGCAAAGGGGCTACAACTGACATCGCATATGATAAAGGA


TCTGTAAAAAAAGATGCCCAACAAGTTCAAAACAAAAAAGTAACCATCCCACAAACAGGTGGTATTGGTACA


ATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCAGTAGTTGTCATGAAAAAACGTCAATCAGAG


GAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 97):


MKKINKCLTVFSTLLLILTSLFSVAPAFADDVTTDTVTLHKIVMPQAAFDNFTEGTKGKNDSDYVGKQINDL


KSYFGSTDAKEIKGAFFVFKNETGTKFITENGKEVDTLEAKDAEGGAVLSGLTKDTGFAFNTAKLKGTYQIV


ELKEKSNYDNNGSILADSKAVPVKITLPLVNNQGVVKDAHIYPKNTETKPQVDKNFADKDLDYTDNRKDKGV


VSATVGDKKEYIVGTKILKGSDYKKLVWTDSMTKGLTFNNNVKVTLDGKDFPVLNYKLVTDDQGFRLALNAT


GLAAVAAAAKDKDVEIKITYSATVNGSTTVEVPETNDVKLDYGNNPTEESEPQEGTPANQEIKVIKDWAVDG


TITDVNVAVKAIFTLQEKQTDGTWVNVASHEATKPSRFEHTFTGLDNTKTYRVVERVSGYTPEYVSFKNGVV


TIKNNKNSNDPTPINPSEPKVVTYGRKFVKTNQANTERLAGATFLVKKEGKYLARKAGAATAEAKAAVKTAK


LALDEAVKAYNDLTKEKQEGQEGKTALATVDQKQKAYNDAFVKANYSYEWVADKKADNVVKLISNAGGQFEI


TGLDKGTYSLEETQAPAGYATLSGDVNFEVTATSYSKGATTDIAYDKGSVKKDAQQVQNKKVTIPQTGGIGT


ILFTIIGLSIMLGAVVVMKKRQSEEA


 


Strain BAA23


ORF DNA sequence (SEQ ID NO: 140):


ATGAAAAAAATCAACAAATGTCTTACAATGTTCTCGACACTGCTATTGATCTTAACGTCACTATTCTCAGTT


GCACCAGCGTTTGCGGACGACGCAACAACTGATACTGTGACCTTGCACAAGATTGTCATGCCACAAGCTGCA


TTTGATAACTTTACTGAAGGTACAAAAGGTAAGAATGATAGCGATTATGTTGGTAAACAAATTAATGACCTT


AAATCTTATTTTGGCTCAACCGATGCTAAAGAAATCAAGGGTGCTTTCTTTGTTTTCAAAAATGAAACTGGT


ACAAAATTCATTACTGAAAATGGTAAGGAAGTCGATACTTTGGAAGCTAAAGATGCTGAAGGTGGTGCTGTT


CTTTCAGGGTTAACAAAAGACAATGGTTTTGTTTTTAACACTGCTAAGTTAAAAGGAATTTACCAAATCGTT


GAATTGAAAGAAAAATCAAACTACGATAACAACGGTTCTATCTTGGCTGATTCAAAAGCAGTTCCAGTTAAA


ATCACTCTGCCATTGGTAAACAACCAAGGTGTTGTTAAAGATGCTCACATTTATCCAAAGAATACTGAAACA


AAACCACAAGTAGATAAGAACTTTGCAGATAAAGATCTTGATTATACTGACAACCGAAAAGACAAAGGTGTT


GTCTCAGCGACAGTTGGTGACAAAAAAGAATACATAGTTGGAACAAAAATTCTTAAAGGCTCAGACTATAAG


AAACTGGTTTGGACTGATAGCATGACTAAAGGTTTGACGTTCAACAACAACGTTAAAGTAACATTGGATGGT


GAAGATTTTCCTGTTTTAAACTACAAACTCGTAACAGATGACCAAGGTTTCCGTCTTGCCTTGAATGCAACA


GGTCTTGCAGCAGTAGCAGCAGCTGCAAAAGACAAAGATGTTGAAATCAAGATCACTTACTCAGCTACGGTG


AACGGCTCCACTACTGTTGAAATTCCAGAAACCAATGATGTTAAATTGGACTATGGTAATAACCCAACGGAA


GAAAGTGAACCACAAGAAGGTACTCCAGCTAACCAAGAAATTAAAGTCATTAAAGACTGGGCAGTAGATGGT


ACAATTACTGATGCTAATGTTGCAGTTAAAGCTATCTTTACCTTGCAAGAAAAACAAACGGATGGTACATGG


GTGAACGTTGCTTCACACGAAGCAACAAAACCATCACGCTTTGAACATACTTTCACAGGTTTGGATAATGCT


AAAACTTACCGCGTTGTCGAACGTGTTAGCGGCTACACTCCAGAATACGTATCATTTAAAAATGGTGTTGTG


ACTATCAAGAACAACAAAAACTCAAATGATCCAACTCCAATCAACCCATCAGAACCAAAAGTGGTGACTTAT


GGACGTAAATTTGTGAAAACAAATCAAGCTAACACTGAACGCTTGGCAGGAGCTACCTTCCTCGTTAAGAAA


GAAGGCAAATACTTGGCACGTAAAGCAGGTGCAGCAACTGCTGAAGCAAAGGCAGCTGTAAAAACTGCTAAA


CTAGCATTGGATGAAGCTGTTAAAGCTTATAACGACTTGACTAAAGAAAAACAAGAAGGCCAAGAAGGTAAA


ACAGCATTGGCTACTGTTGATCAAAAACAAAAAGCTTACAATGACGCTTTTGTTAAAGCTAACTACTCATAT


GAATGGGTTGCAGATAAAAAGGCTGATAATGTTGTTAAATTGATCTCTAACGCCGGTGGTCAATTTGAAATT


ACTGGTTTGGATAAAGGCACTTATGGCTTGGAAGAAACTCAAGCACCAGCAGGTTATGCGACATTGTCAGGT


GATGTAAACTTTGAAGTAACTGCCACATCATATAGCAAAGGGGCTACAACTGACATCGCATATGATAAAGGC


TCTGTAAAAAAAGATGCCCAACAAGTTCAAAACAAAAAAGTAACCATCCCACAAACAGGTGGTATTGGTACA


ATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCAGTAGTTATCATGAAAAAACGTCAATCAGAG


GAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 98):


MKKINKCLTMFSTLLLILTSLFSVAPAFADDATTDTVTLHKIVMPQAAFDNFTEGTKGKNDSDYVGKQINDL


KSYFGSTDAKEIKGAFFVFKNETGTKFITENGKEVDTLEAKDAEGGAVLSGLTKDNGFVFNTAKLKGIYQIV


ELKEKSNYDNNGSILADSKAVPVKITLPLVNNQGVVKDAHIYPKNTETKPQVDKNFADKDLDYTDNRKDKGV


VSATVGDKKEYIVGTKILKGSDYKKLVWTDSMTKGLTFNNNVKVTLDGEDFPVLNYKLVTDDQGFRLALNAT


GLAAVAAAAKDKDVEIKITYSATVNGSTTVEIPETNDVKLDYGNNPTEESEPQEGTPANQEIKVIKDWAVDG


TITDANVAVKAIFTLQEKQTDGTWVNVASHEATKPSRFEHTFTGLDNAKTYRVVERVSGYTPEYVSFKNGVV


TIKNNKNSNDPTPINPSEPKVVTYGRKFVKTNQANTERLAGATFLVKKEGKYLARKAGAATAEAKAAVKTAK


LALDEAVKAYNDLTKEKQEGQEGKTALATVDQKQKAYNDAFVKANYSYEWVADKKADNVVKLISNAGGQFEI


TGLDKGTYGLEETQAPAGYATLSGDVNFEVTATSYSKGATTDIAYDKGSVKKDAQQVQNKKVTIPQTGGIGT


ILFTIIGLSIMLGAVVIMKKRQSEEA


 


Strain BAA611


ORF DNA sequence (SEQ ID NO: 141):


ATGAAAAAAATCAACAAATGTCTTACAATGTTCTCGACACTGCTATTGATCTTAACGTCACTATTCTCAGTT


GCACCAGCGTTTGCGGACGACGCAACAACTGATACTGTGACCTTGCACAAGATTGTCATGCCACAAGCTGCA


TTTGATAACTTTACTGAAGGTACAAAAGGTAAGAATGATAGCGATTATGTTGGTAAACAAATTAATGACCTT


AAATCTTATTTTGGCTCAACCGATGCTAAAGAAATCAAGGGTGCTTTCTTTGTTTTCAAAAATGAAACTGGT


ACAAAATTCATTACTGAAAATGGTAAGGAAGTCGATACTTTGGAAGCTAAAGATGCTGAAGGTGGTGCTGTT


CTTTCAGGGTTAACAAAAGACAATGGTTTTGTTTTTAACACTGCTAAGTTAAAAGGAATTTACCAAATCGTT


GAATTGAAAGAAAAATCAAACTACGATAACAACGGTTCTATCTTGGCTGATTCAAAAGCAGTTCCAGTTAAA


ATCACTCTGCCATTGGTAAACAACCAAGGTGTTGTTAAAGATGCTCACATTTATCCAAAGAATACTGAAACA


AAACCACAAGTAGATAAGAACTTTGCAGATAAAGATCTTGATTATACTGACAACCGAAAAGACAAAGGTGTT


GTCTCAGCGACAGTTGGTGACAAAAAAGAATACATAGTTGGAACAAAAATTCTTAAAGGCTCAGACTATAAG


AAACTGGTTTGGACTGATAGCATGACTAAAGGTTTGACGTTCAACAACAACGTTAAAGTAACATTGGATGGT


GAAGATTTTCCTGTTTTAAACTACAAACTCGTAACAGATGACCAAGGTTTCCGTCTTGCCTTGAATGCAACA


GGTCTTGCAGCAGTAGCAGCAGCTGCAAAAGACAAAGATGTTGAAATCAAGATCACTTACTCAGCTACGGTG


AACGGCTCCACTACTGTTGAAATTCCAGAAACCAATGATGTTAAATTGGACTATGGTAATAACCCAACGGAA


GAAAGTGAACCACAAGAAGGTACTCCAGCTAACCAAGAAATTAAAGTCATTAAAGACTGGGCAGTAGATGGT


ACAATTACTGATGCTAATGTTGCAGTTAAAGCTATCTTTACCTTGCAAGAAAAACAAACGGATGGTACATGG


GTGAACGTTGCTTCACACGAAGCAACAAAACCATCACGCTTTGAACATACTTTCACAGGTTTGGATAATGCT


AAAACTTACCGCGTTGTCGAACGTGTTAGCGGCTACACTCCAGAATACGTATCATTTAAAAATGGTGTTGTG


ACTATCAAGAACAACAAAAACTCAAATGATCCAACTCCAATCAACCCATCAGAACCAAAAGTGGTGACTTAT


GGACGTAAATTTGTGAAAACAAATCAAGCTAACACTGAACGCTTGGCAGGAGCTACCTTCCTCGTTAAGAAA


GAAGGCAAATACTTGGCACGTAAAGCAGGTGCAGCAACTGCTGAAGCAAAGGCAGCTGTAAAAACTGCTAAA


CTAGCATTGGATGAAGCTGTTAAAGCTTATAACGACTTGACTAAAGAAAAACAAGAAGGCCAAGAAGGTAAA


ACAGCATTGGCTACTGTTGATCAAAAACAAAAAGCTTACAATGACGCTTTTGTTAAAGCTAACTACTCATAT


GAATGGGTTGCAGATAAAAAGGCTGATAATGTTGTTAAATTGATCTCTAACGCCGGTGGTCAATTTGAAATT


ACTGGTTTGGATAAAGGCACTTATGGCTTGGAAGAAACTCAAGCACCAGCAGGTTATGCGACATTGTCAGGT


GATGTAAACTTTGAAGTAACTGCCACATCATATAGCAAAGGGGCTACAACTGACATCGCATATGATAAAGGC


TCTGTAAAAAAAGATGCCCAACAAGTTCAAAACAAAAAAGTAACCATCCCACAAACAGGTGGTATTGGTACA


ATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCAGTAGTTATCATGAAAAAACGTCAATCAGAG


GAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 99):


MKKINKCLTMFSTLLLILTSLFSVAPAFADDATTDTVTLHKIVMPQAAFDNFTEGTKGKNDSDYVGKQINDL


KSYFGSTDAKEIKGAFFVFKNETGTKFITENGKEVDTLEAKDAEGGAVLSGLTKDNGFVFNTAKLKGIYQIV


ELKEKSNYDNNGSILADSKAVPVKITLPLVNNQGVVKDAHIYPKNTETKPQVDKNFADKDLDYTDNRKDKGV


VSATVGDKKEYIVGTKILKGSDYKKLVWTDSMTKGLTFNNNVKVTLDGEDFPVLNYKLVTDDQGFRLALNAT


GLAAVAAAAKDKDVEIKITYSATVNGSTTVEIPETNDVKLDYGNNPTEESEPQEGTPANQEIKVIKDWAVDG


TITDANVAVKAIFTLQEKQTDGTWVNVASHEATKPSRFEHTFTGLDNAKTYRVVERVSGYTPEYVSFKNGVV


TIKNNKNSNDPTPINPSEPKVVTYGRKFVKTNQANTERLAGATFLVKKEGKYLARKAGAATAEAKAAVKTAK


LALDEAVKAYNDLTKEKQEGQEGKTALATVDQKQKAYNDAFVKANYSYEWVADKKADNVVKLISNAGGQFEI


TGLDKGTYGLEETQAPAGYATLSGDVNFEVTATSYSKGATTDIAYDKGSVKKDAQQVQNKKVTIPQTGGIGT


ILFTIIGLSIMLGAVVIMKKRQSEEA


 


Strain C388/90


ORF DNA sequence (SEQ ID NO: 142):


ATGAAAAAAATCAACAAATATTTTGCAGTCTTCTCGGCATTGCTACTGACCGTAACATCATTGTTCTCAGTT


GCACCAGTGTTTGCGGAAGAAGCAAAAACTACTGACACAGTGACCTTGCACAAGATTGTCATGCCTCGAACT


GCATTTGACGGTTTTACTGCTGGTACAAAGGGTAAGGATAATACTGACTACGTTGGTAAACAAATCGAAGAC


CTTAAAACTTACTTTGGCTCAGGCGAAGCGAAAGAAATCGCAGGTGCTTACTTTGCTTTCAAAAATGAAGCT


GGTACTAAATACATCACTGAAAATGGTGAAGAAGTTGATACTTTGGATACAACAGATGCCAAAGGTGGTGCT


GTTCTTAAAGGTTTAACAACAGACAATGGTTTCAAATTTAACACTTCTAAATTAACAGGAACTTACCAAATC


GTTGAATTGAAAGAAAAATCTACATACAACAACGATGGTTCTATCTTGGCTGATTCAAAAGCAGTTCCAGTT


AAAATCACTCTTCCATTGGTAAACGACAATGGTGTTGTTAAAGACGCTCACGTTTATCCAAAGAACACTGAA


ACAAAACCACAAGTAGATAAGAACTTCGCAGATAAAGAACTTGATTATGCGAACAACAAAAAAGACAAAGGG


ACTGTCTCAGCATCTGTTGGTGATGTTAAAAAATATCATGTTGGAACAAAAATCCTTAAAGGTTCAGACTAT


AAGAAATTAATCTGGACCGATAGCATGACCAAAGGTTTGACTTTCAACAACGATATTGCTGTAACATTGGAT


GGTGCAACTCTTGATGCTACAAATTACAAACTTGTAGCAGATGACCAAGGTTTCCGCCTTGTCTTGACTGAC


AAAGGTCTTGAAGCAGTGGCAAAAGCCGCAAAAACAAAAGATGTTGAAATCAAGATCACTTACTCAGCTACT


TTGAACGGTTCTGCTGTCGTTGAAGTTCTAGAAACCAATGATGTTAAATTGGACTACGGCAACAACCCAACA


ATTGAAAATGAACCAAAAGAAGGTATTCCAGTTGATAAGAAAATCACTGTTAACAAAACATGGGCAGTAGAT


GGCAATGAAGTGAATAAAGCAGATGAAACAGTTGATGCTGTCTTCACCTTGCAAGTTAAAGATGGTGACAAA


TGGGTGAATGTTGATTCAGCTAAAGCAACAGCTGCAACTAGCTTCAAACACACTTTTGAAAACTTGGATAAT


GCTAAAACTTACCGCGTTATCGAACGTGTTAGCGGCTACGCTCCAGAATACGTCTCATTTGTAAATGGCGTT


GTAACCATCAAGAACAACAAAGACTCAAATGAGCCAACTCCAATCAACCCATCAGAACCAAAAGTGGTGACT


TATGGACGTAAATTTGTGAAAACAAATAAAGATGGAAAAGAACGCTTGGCAGGAGCTACCTTCCTTGTTAAG


AAAGATGGCAAGTACTTGGCACGTAAATCAGGTGTTGCAACAGATGCAGAAAAAGCTGCTGTAGATTCAACT


AAATCAGCATTGGATGCTGCTGTTAAAGCTTACAATGATTTGACTAAAGAAAAACAAGAAGGTCAAGATGGT


AAATCAGCATTGGCTACCGTTAGTGAAAAACAAAAAGCTTACAATGATGCCTTTGTTAAAGCTAACTACTCA


TACGAATGGGTTGAAGATAAAAATGCTAAGAATGTTGTTAAATTGATTTCTAACGATAAAGGTCAATTTGAA


ATTACTGGCTTGACTGAAGGTCAATACTCATTGGAAGAAACACAAGCACCAACTGGTTATGCTAAATTATCA


GGTGATGTTTCGTTTAATGTTAATGCTACTTCATACAGTAAAGGTTCTGCTCAAGATATTGAGTATACCCAA


GGTTCTAAAACTAAAGATGCACAACAAGTTATCAATAAGAAGGTTACTATTCCACAAACAGGTGGTATTGGT


ACAATTTTTTTCACAATTATTGGATTAAGTATTATGCTTGGAGCGGTAGTTATCATGAAAAGACGTCAATCA


GAGGAAGTTTAA


 


ORF amino acid sequence (SEQ ID NO: 100):


MKKINKYFAVFSALLLTVTSLFSVAPVFAEEAKTTDTVTLHKIVMPRTAFDGFTAGTKGKDNTDYVGKQIED


LKTYFGSGEAKEIAGAYFAFKNEAGTKYITENGEEVDTLDTTDAKGGAVLKGLTTDNGFKFNTSKLTGTYQI


VELKEKSTYNNDGSILADSKAVPVKITLPLVNDNGVVKDAHVYPKNTETKPQVDKNFADKELDYANNKKDKG


TVSASVGDVKKYHVGTKILKGSDYKKLIWTDSMTKGLTFNNDIAVTLDGATLDATNYKLVADDQGFRLVLTD


KGLEAVAKAAKTKDVEIKITYSATLNGSAVVEVLETNDVKLDYGNNPTIENEPKEGIPVDKKITVNKTWAVD


GNEVNKADETVDAVFTLQVKDGDKWVNVDSAKATAATSFKHTFENLDNAKTYRVIERVSGYAPEYVSFVNGV


VTIKNNKDSNEPTPINPSEPKVVTYGRKFVKTNKDGKERLAGATFLVKKDGKYLARKSGVATDAEKAAVDST


KSALDAAVKAYNDLTKEKQEGQDGKSALATVSEKQKAYNDAFVKANYSYEWVEDKNAKNVVKLISNDKGQFE


ITGLTEGQYSLEETQAPTGYAKLSGDVSFNVNATSYSKGSAQDIEYTQGSKTKDAQQVINKKVTIPQTGGIG


TIFFTIIGLSIMLGAVVIMKRRQSEEV


 


Strain IC97


ORF DNA sequence (SEQ ID NO: 143):


ATGAAAAGAATCAACAAATATTTTGCAATGTTCTCGGCATTGTTATTAATTTTAACATCGTTGTTATCGGTA


GCTCCGGTATTTGCTGCTGAGATGGGAAATATCACTAAAACAGTAACCTTACACAAAATTGTTCAAACATCC


GATAATTTGGCTAAGCCAAATTTCCCAGGAATAAATGGATTGAATGGAACGAAGTATATGGGTCAAAAACTT


ACTGACATTTCAGGATATTTTGGGCAAGGTTCTAAAGAAATCGCCGGTGCTTTCTTTGCGGTTATGAATGAA


AGTCAGACAAAATATATCACAGAAAGTGGTACTGAAGTAGAAAGTATCGATGCAGCAGGTGTCCTTAAAGGT


TTGACAACTGAAAACGGCATTACATTTAATACTGCAAACTTAAAAGGAACATACCAAATCGTTGAGTTGCTT


GACAAATCTAATTATAAAAATGGTGACAAAGTTCTTGCTGACTCAAAAGCTGTCCCAGTGAAAATCACTCTT


CCTTTGTATAACGAAGAAGGAATTGTCGTGGACGCTGAAGTGTATCCAAAGAATACAGAAGAAGCACCACAA


ATCGACAAAAACTTTGCTAAAGCAAATAAATTGTTGAATGACAGTGATAATTCAGCTATTGCAGGTGGGGCA


GACTACGACAAATATCAGGCAGAAAAAGCAAAAGCTACTGCTGAAATCGGTCAAGAAATCCCTTACGAAGTT


AAAACAAAAATCCAAAAAGGGTCTAAATACAAAAACCTTGCTTGGGTCGATACCATGTCAAATGGTTTGACA


ATGGGTAACACTGTTAACTTAGAAGCATCGTCAGGCTCTTTTGTAGAAGGTACAGATTACAATGTTGAACGT


GATGACCGTGGTTTCACTTTGAAATTCACAGATACAGGTTTGACTAAGCTACAAAAAGAAGCGGAAACACAA


GCTGTTGAATTCACATTGACATATAGCGCAACAGTTAACGGTGCGGCTATTGATGACAAGCCAGAAAGCAAT


GATATCAAACTTCAATACGGTAACAAACCAGGTAAAAAAGTAAAAGAAATCCCAGTAACACCGTCAAATGGC


GAAATCACTGTTAGCAAAACTTGGGACAAAGGTTCAGATTTAGAGAATGCGAATGTTGTTTATACCCTTAAA


GATGGTGGAACAGCTGTTGCCTCAGTTTCATTGACAAAAACAACACCAAATGGCGAAATCAACTTAGGTAAT


GGTATTAAATTTACAGTTACTGGAGCGTTTGCTGGTAAATTCAGTGGTCTGACTGATAGTAAAACATACATG


ATCTCAGAACGTATCGCTGGTTATGGTAATACAATCACTACTGGTGCTGGTAGTGCAGCTATCACCAATACT


CCAGATTCAGACAACCCAACACCACTTAATCCAACTGAACCAAAAGTTGTGACACACGGTAAAAAATTCGTC


AAAACAAGTTCGACTGAAACAGAACGCTTGCAAGGTGCACAGTTCGTTGTTAAAGATTCAGCTGGTAAATAC


CTTGCATTGAAATCATCTGCGACAATATCAGCTCAAACAACAGCTTACACAAATGCTAAAACTGCTCTTGAC


GCTAAAATCGCAGCTTACAACAAACTTTCAGCAGACGATCAAAAAGGTACTAAAGGTGAAACAGCTAAAGCA


GAAATCAAAACTGCTCAAGACGCTTACAATGCAGCCTTCATCGTAGCTCGTACAGCTTACGAGTGGGTAACT


AATAAAGAAGATGCTAACGTTGTTAAAGTGACTTCAAACGCTGACGGTCAATTTGAAGTTAGCGGTCTTGCA


ACTGGTGATTATAAACTTGAAGAAACACAAGCTCCAGCTGGTTACGCTAAATTAGCAGGTGATGTTGATTTC


AAAGTTGGAAACAGCTCAAAAGCAGACGACTCAGGTAACATTGATTACACTGCTAGCAGCAATAAAAAAGAC


GCTCAACGCATAGAAAACAAAAAAGTGACTATTCCACAAACAGGTGGTATTGGTACAATTCTTTTCACAATT


ATTGGTTTAAGCATTATGCTTGGAGCGGTAATTATCATGAAAAGACGTCAATCAGAGGAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 101):


MKRINKYFAMFSALLLILTSLLSVAPVFAAEMGNITKTVTLHKIVQTSDNLAKPNFPGINGLNGTKYMGQKL


TDISGYFGQGSKEIAGAFFAVMNESQTKYITESGTEVESIDAAGVLKGLTTENGITFNTANLKGTYQIVELL


DKSNYKNGDKVLADSKAVPVKITLPLYNEEGIVVDAEVYPKNTEEAPQIDKNFAKANKLLNDSDNSAIAGGA


DYDKYQAEKAKATAEIGQEIPYEVKTKIQKGSKYKNLAWVDTMSNGLTMGNTVNLEASSGSFVEGTDYNVER


DDRGFTLKFTDTGLTKLQKEAETQAVEFTLTYSATVNGAAIDDKPESNDIKLQYGNKPGKKVKEIPVTPSNG


EITVSKTWDKGSDLENANVVYTLKDGGTAVASVSLTKTTPNGEINLGNGIKFTVTGAFAGKFSGLTDSKTYM


ISERIAGYGNTITTGAGSAAITNTPDSDNPTPLNPTEPKVVTHGKKFVKTSSTETERLQGAQFVVKDSAGKY


LALKSSATISAQTTAYTNAKTALDAKIAAYNKLSADDQKGTKGETAKAEIKTAQDAYNAAFIVARTAYEWVT


NKEDANVVKVTSNADGQFEVSGLATGDYKLEETQAPAGYAKLAGDVDFKVGNSSKADDSGNIDYTASSNKKD


AQRIENKKVTIPQTGGIGTILFTIIGLSIMLGAVIIMKRRQSEEA


 


Strain IC98


ORF DNA sequence (SEQ ID NO: 144):


ATGAAAAAAATCAACAAATGTCTTACAATGTTCTCGACACTGCTATTGATCTTAACGTCACTATTCTCAGTT


GCACCAGCGTTTGCGGACGACGCAACAACTGATACTGTGACCTTGCACAAGATTGTCATGCCACAAGCTGCA


TTTGATAACTTTACTGAAGGTACAAAAGGTAAGAATGATAGCGATTATGTTGGTAAACAAATTAATGACCTT


AAATCTTATTTTGGCTCAACCGATGCTAAAGAAATCAAGGGTGCTTTCTTTGTTTTCAAAAATGAAACTGGT


ACAAAATTCATTACTGAAAATGGTAAGGAAGTCGATACTTTGGAAGCTAAAGATGCTGAAGGTGGTGCTGTT


CTTTCAGGGTTAACAAAAGACAATGGTTTTGTTTTTAACACTGCTAAGTTAAAAGGAATTTACCAAATCGTT


GAATTGAAAGAAAAATCAAACTACGATAACAACGGTTCTATCTTGGCTGATTCAAAAGCAGTTCCAGTTAAA


ATCACTCTGCCATTGGTAAACAACCAAGGTGTTGTTAAAGATGCTCACATTTATCCAAAGAATACTGAAACA


AAACCACAAGTAGATAAGAACTTTGCAGATAAAGATCTTGATTATACTGACAACCGAAAAGACAAAGGTGTT


GTCTCAGCGACAGTTGGTGACAAAAAAGAATACATAGTTGGAACAAAAATTCTTAAAGGCTCAGACTATAAG


AAACTGGTTTGGACTGATAGCATGACTAAAGGTTTGACGTTCAACAACAACGTTAAAGTAACATTGGATGGT


GAAGATTTTCCTGTTTTAAACTACAAACTCGTAACAGATGACCAAGGTTTCCGTCTTGCCTTGAATGCAACA


GGTCTTGCAGCAGTAGCAGCAGCTGCAAAAGACAAAGATGTTGAAATCAAGATCACTTACTCAGCTACGGTG


AACGGCTCCACTACTGTTGAAATTCCAGAAACCAATGATGTTAAATTGGACTATGGTAATAACCCAACGGAA


GAAAGTGAACCACAAGAAGGTACTCCAGCTAACCAAGAAATTAAAGTCATTAAAGACTGGGCAGTAGATGGT


ACAATTACTGATGCTAATGTTGCAGTTAAAGCTATCTTTACCTTGCAAGAAAAACAAACGGATGGTACATGG


GTGAACGTTGCTTCACACGAAGCAACAAAACCATCACGCTTTGAACATACTTTCACAGGTTTGGATAATGCT


AAAACTTACCGCGTTGTCGAACGTGTTAGCGGCTACACTCCAGAATACGTATCATTTAAAAATGGTGTTGTG


ACTATCAAGAACAACAAAAACTCAAATGATCCAACTCCAATCAACCCATCAGAACCAAAAGTGGTGACTTAT


GGACGTAAATTTGTGAAAACAAATCAAGCTAACACTGAACGCTTGGCAGGAGCTACCTTCCTCGTTAAGAAA


GAAGGCAAATACTTGGCACGTAAAGCAGGTGCAGCAACTGCTGAAGCAAAGGCAGCTGTAAAAACTGCTAAA


CTAGCATTGGATGAAGCTGTTAAAGCTTATAACGACTTGACTAAAGAAAAACAAGAAGGCCAAGAAGGTAAA


ACAGCATTGGCTACTGTTGATCAAAAACAAAAAGCTTACAATGACGCTTTTGTTAAAGCTAACTACTCATAT


GAATGGGTTGCAGATAAAAAGGCTGATAATGTTGTTAAATTGATCTCTAACGCCGGTGGTCAATTTGAAATT


ACTGGTTTGGATAAAGGCACTTATGGCTTGGAAGAAACTCAAGCACCAGCAGGTTATGCGACATTGTCAGGT


GATGTAAACTTTGAAGTAACTGCCACATCATATAGCAAAGGGGCTACAACTGACATCGCATATGATAAAGGC


TCTGTAAAAAAAGATGCCCAACAAGTTCAAAACAAAAAAGTAACCATCCCACAAACAGGTGGTATTGGTACA


ATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCAGTAGTTATCATGAAAAAACGTCAATCAGAG


GAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 102):


MKKINKCLTMFSTLLLILTSLFSVAPAFADDATTDTVTLHKIVMPQAAFDNFTEGTKGKNDSDYVGKQINDL


KSYFGSTDAKEIKGAFFVFKNETGTKFITENGKEVDTLEAKDAEGGAVLSGLTKDNGFVFNTAKLKGIYQIV


ELKEKSNYDNNGSILADSKAVPVKITLPLVNNQGVVKDAHIYPKNTETKPQVDKNFADKDLDYTDNRKDKGV


VSATVGDKKEYIVGTKILKGSDYKKLVWTDSMTKGLTFNNNVKVTLDGEDFPVLNYKLVTDDQGFRLALNAT


GLAAVAAAAKDKDVEIKITYSATVNGSTTVEIPETNDVKLDYGNNPTEESEPQEGTPANQEIKVIKDWAVDG


TITDANVAVKAIFTLQEKQTDGTWVNVASHEATKPSRFEHTFTGLDNAKTYRVVERVSGYTPEYVSFKNGVV


TIKNNKNSNDPTPINPSEPKVVTYGRKFVKTNQANTERLAGATFLVKKEGKYLARKAGAATAEAKAAVKTAK


LALDEAVKAYNDLTKEKQEGQEGKTALATVDQKQKAYNDAFVKANYSYEWVADKKADNVVKLISNAGGQFEI


TGLDKGTYGLEETQAPAGYATLSGDVNFEVTATSYSKGATTDIAYDKGSVKKDAQQVQNKKVTIPQTGGIGT


ILFTIIGLSIMLGAVVIMKKRQSEEA


 


Strain IC105


ORF DNA sequence (SEQ ID NO: 145):


ATGAAAAGAATCAACAAATATTTTGCAATGTTCTCGGCATTGTTATTAATTTTAACATCGTTGTTATCGGTA


GCTCCGGTATTTGCTGCTGAGATGGGAAATATCACTAAAACAGTAACCTTACACAAAATTGTTCAAACATCC


GATAATTTGGCTAAGCCAAATTTCCCAGGAATAAATGGATTGAATGGAACGAAGTATATGGGTCAAAAACTT


ACTGACATTTCAGGATATTTTGGGCAAGGTTCTAAAGAAATCGCCGGTGCTTTCTTTGCGGTTATGAATGAA


AGTCAGACAAAATATATCACAGAAAGTGGTACTGAAGTAGAAAGTATCGATGCAGCAGGTGTCCTTAAAGGT


TTGACAACTGAAAACGGCATTACATTTAATACTGCAAACTTAAAAGGAACATACCAAATCGTTGAGTTGCTT


GACAAATCTAATTATAAAAATGGTGACAAAGTTCTTGCTGACTCAAAAGCTGTCCCAGTGAAAATCACTCTT


CCTTTGTATAACGAAGAAGGAATTGTCGTGGACGCTGAAGTGTATCCAAAGAATACAGAAGAAGCACCACAA


ATCGACAAAAACTTTGCTAAAGCAAATAAATTGTTGAATGACAGTGATAATTCAGCTATTGCAGGTGGGGCA


GACTACGACAAATATCAGGCAGAAAAAGCAAAAGCTACTGCTGAAATCGGTCAAGAAATCCCTTACGAAGTT


AAAACAAAAATCCAAAAAGGGTCTAAATACAAAAACCTTGCTTGGGTCGATACCATGTCAAATGGTTTGACA


ATGGGTAACACTGTTAACTTAGAAGCATCGTCAGGCTCTTTTGTAGAAGGTACAGATTACAATGTTGAACGT


GATGACCGTGGTTTCACTTTGAAATTCACAGATACAGGTTTGACTAAGCTACAAAAAGAAGCGGAAACACAC


GCTGTTGAATTCACATTGACATATAGCGCAACAGTTAACGGTGCGGCTATTGATGACAAGCCAGAAAGCAAT


GATATCAAACTTCAATACGGTAACAAACCAGGTAAAAAAGTAAAAGAAATCCCAGTAACACCGTCAAATGGC


GAAATCACTGTTAGCAAAACTTGGGACAAAGGTTCAGATTTAGAGAATGCGAATGTTGTTTATACCCTTAAA


GATGGTGGAACAGCTGTTGCCTCAGTTTCATTGACAAAAACAACACCAAATGGCGAAATCAACTTAGGTAAT


GGTATTAAATTTACAGTTACTGGAGCGTTTGCTGGTAAATTCAGTGGTCTGACTGATAGTAAAACATACATG


ATCTCAGAACGTATCGCTGGTTATGGTAATACAATCACTACTGGTGCTGGTAGTGCAGCTATCACCAATACT


CCAGATTCAGACAACCCAACACCACTTAATCCAACTGAACCAAAAGTTGTGACACACGGTAAAAAATTCGTC


AAAACAAGTTCGACTGAAACAGAACGCTTGCAAGGTGCACAGTTCGTTGTTAAAGATTCAGCTGGTAAATAC


CTTGCATTGAAATCATCTGCGACAATATCAGCTCAAACAACAGCTTACACAAATGCTAAAACTGCTCTTGAC


GCTAAAATCGCAGCTTACAACAAACTTTCAGCAGACGATCAAAAAGGTACTAAAGGTGAAACAGCTAAAGCA


GAAATCAAAACTGCTCAAGACGCTTACAATGCAGCCTTCATCGTAGCTCGTACAGCTTACGAGTGGGTAACT


AATAAAGAAGATGCTAACGTTGTTAAAGTGACTTCAAACGCTGACGGTCAATTTGAAGTTAGCGGTCTTGCA


ACTGGTGATTATAAACTTGAAGAAACACAAGCTCCAGCTGGTTACGCTAAATTAGCAGGTGATGTTGATTTC


AAAGTTGGAAACAGCTCAAAAGCAGACGACTCAGGTAACATTGATTACACTGCTAGCAGCAATAAAAAAGAC


GCTCAACGCATAGAAAACAAAAAAGTGACTATTCCACAAACAGGTGGTATTGGTACAATTCTTTTCACAATT


ATTGGTTTAAGCATTATGCTTGGAGCGGTAATTATCATGAAAAGACGTCAATCAGAGGAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 103):


MKRINKYFAMFSALLLILTSLLSVAPVFAAEMGNITKTVTLHKIVQTSDNLAKPNFPGINGLNGTKYMGQKL


TDISGYFGQGSKEIAGAFFAVMNESQTKYITESGTEVESIDAAGVLKGLTTENGITFNTANLKGTYQIVELL


DKSNYKNGDKVLADSKAVPVKITLPLYNEEGIVVDAEVYPKNTEEAPQIDKNFAKANKLLNDSDNSAIAGGA


DYDKYQAEKAKATAEIGQEIPYEVKTKIQKGSKYKNLAWVDTMSNGLTMGNTVNLEASSGSFVEGTDYNVER


DDRGFTLKFTDTGLTKLQKEAETHAVEFTLTYSATVNGAAIDDKPESNDIKLQYGNKPGKKVKEIPVTPSNG


EITVSKTWDKGSDLENANVVYTLKDGGTAVASVSLTKTTPNGEINLGNGIKFTVTGAFAGKFSGLTDSKTYM


ISERIAGYGNTITTGAGSAAITNTPDSDNPTPLNPTEPKVVTHGKKFVKTSSTETERLQGAQFVVKDSAGKY


LALKSSATISAQTTAYTNAKTALDAKIAAYNKLSADDQKGTKGETAKAEIKTAQDAYNAAFIVARTAYEWVT


NKEDANVVKVTSNADGQFEVSGLATGDYKLEETQAPAGYAKLAGDVDFKVGNSSKADDSGNIDYTASSNKKD


AQRIENKKVTIPQTGGIGTILFTIIGLSIMLGAVIIMKRRQSEEA


 


Strain IC216


ORF DNA sequence (SEQ ID NO: 146):


ATGAAAAAAATCAACAAATGTCTTACAATGTTCTCGACACTGCTATTGATCTTAACGTCACTATTCTCAGTT


GCACCAGCGTTTGCGGACGACGCAACAACTGATACTGTGACCTTGCACAAGATTGTCATGCCACAAGCTGCA


TTTGATAACTTTACTGAAGGTACAAAAGGTAAGAATGATAGCGATTATGTTGGTAAACAAATTAATGACCTT


AAATCTTATTTTGGCTCAACCGATGCTAAAGAAATCAAGGGTGCTTTCTTTGTTTTCAAAAATGAAACTGGT


ACAAAATTCATTACTGAAAATGGTAAGGAAGTCGATACTTTGGAAGCTAAAGATGCTGAAGGTGGTGCTGTT


CTTTCAGGGTTAACAAAAGACAATGGTTTTGTTTTTAACACTGCTAAGTTAAAAGGAATTTACCAAATCGTT


GAATTGAAAGAAAAATCAAACTACGATAACAACGGTTCTATCTTGGCTGATTCAAAAGCAGTTCCAGTTAAA


ATCACTCTGCCATTGGTAAACAACCAAGGTGTTGTTAAAGATGCTCACATTTATCCAAAGAATACTGAAACA


AAACCACAAGTAGATAAGAACTTTGCAGATAAAGATCTTGATTATACTGACAACCGAAAAGACAAAGGTGTT


GTCTCAGCGACAGTTGGTGACAAAAAAGAATACATAGTTGTAACAAAAATTCTTAAAGGCTCAGACTATAAG


AAACTGGTTTGGACTGATAGCATGACTAAAGGTTTGACGTTCAACAACAACGTTAAAGTAACATTGGATGGT


GAAGATTTTCCTGTTTTAAACTACAAACTCGTAACAGATGACCAAGGTTTCCGTCTTGCCTTGAATGCAACA


GGTCTTGCAGCAGTAGCAGCAGCTGCAAAAGACAAAGATGTTGAAATCAAGATCACTTACTCAGCTACGGTG


AACGGCTCCACTACTGTTGAAATTCCAGAAACCAATGATGTTAAATTGGACTATGGTAATAACCCAACGGAA


GAAAGTGAACCACAAGAAGGTACTCCAGCTAACCAAGAAATTAAAGTCATTAAAGACTGGGCAGTAGATGGT


ACAATTACTGATGCTAATGTTGCAGTTAAAGCTATCTTTACCTTGCAAGAAAAACAAACGGATGGTACATGG


GTGAACGTTGCTTCACACGAAGCAACAAAACCATCACGCTTTGAACATACTTTCACAGGTTTGGATAATGCT


AAAACTTACCGCGTTGTCGAACGTGTTAGCGGCTACACTCCAGAATACGTATCATTTAAAAATGGTGTTGTG


ACTATCAAGAACAACAAAAACTCAAATGATCCAACTCCAATCAACCCATCAGAACCAAAAGTGGTGACTTAT


GGACGTAAATTTGTGAAAACAAATCAAGCTAACACTGAACGCTTGGCAGGAGCTACCTTCCTCGTTAAGAAA


GAAGGCAAATACTTGGCACGTAAAGCAGGTGCAGCAACTGCTGAAGCAAAGGCAGCTGTAAAAACTGCTAAA


CTAGCATTGGATGAAGCTGTTAAAGCTTATAACGACTTGACTAAAGAAAAACAAGAAGGCCAAGAAGGTAAA


ACAGCATTGGCTACTGTTGATCAAAAACAAAAAGCTTACAATGACGCTTTTGTTAAAGCTAACTACTCATAT


GAATGGGTTGCAGATAAAAAGGCTGATAATGTTGTTAAATTGATCTCTAACGCCGGTGGTCAATTTGAAATT


ACTGGTTTGGATAAAGGCACTTATGGCTTGGAAGAAACTCAAGCACCAGCAGGTTATGCGACATTGTCAGGT


GATGTAAACTTTGAAGTAACTGCCACATCATATAGCAAAGGGGCTACAACTGACATCGCATATGATAAAGGC


TCTGTAAAAAAAGATGCCCAACAAGTTCAAAACAAAAAAGTAACCATCCCACAAACAGGTGGTATTGGTACA


ATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCAGTAGTTATCATGAAAAAACGTCAATCAGAG


GAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 104):


MKKINKCLTMFSTLLLILTSLFSVAPAFADDATTDTVTLHKIVMPQAAFDNFTEGTKGKNDSDYVGKQINDL


KSYFGSTDAKEIKGAFFVFKNETGTKFITENGKEVDTLEAKDAEGGAVLSGLTKDNGFVFNTAKLKGIYQIV


ELKEKSNYDNNGSILADSKAVPVKITLPLVNNQGVVKDAHIYPKNTETKPQVDKNFADKDLDYTDNRKDKGV


VSATVGDKKEYIVVTKILKGSDYKKLVWTDSMTKGLTFNNNVKVTLDGEDFPVLNYKLVTDDQGFRLALNAT


GLAAVAAAAKDKDVEIKITYSATVNGSTTVEIPETNDVKLDYGNNPTEESEPQEGTPANQEIKVIKDWAVDG


TITDANVAVKAIFTLQEKQTDGTWVNVASHEATKPSRFEHTFTGLDNAKTYRVVERVSGYTPEYVSFKNGVV


TIKNNKNSNDPTPINPSEPKVVTYGRKFVKTNQANTERLAGATFLVKKEGKYLARKAGAATAEAKAAVKTAK


LALDEAVKAYNDLTKEKQEGQEGKTALATVDQKQKAYNDAFVKANYSYEWVADKKADNVVKLISNAGGQFEI


TGLDKGTYGLEETQAPAGYATLSGDVNFEVTATSYSKGATTDIAYDKGSVKKDAQQVQNKKVTIPQTGGIGT


ILFTIIGLSIMLGAVVIMKKRQSEEA


 


Strain IC245


ORF DNA sequence (SEQ ID NO: 147):


ATGAAAAAAATCAACAAATGTCTTACAATGTTCTCGACACTGCTATTGATCTTAACGTCACTATTCTCAGTT


GCACCAGCGTTTGCGGACGACGCAACAACTGATACTGTGACCTTGCACAAGATTGTCATGCCACAAGCTGCA


TTTGATAACTTTACTGAAGGTACAAAAGGTAAGAATGATAGCGATTATGTTGGTAAACAAATTAATGACCTT


AAATCTTATTTTGGCTCAACCGATGCTAAAGAAATCAAGGGTGCTTTCTTTGTTTTCAAAAATGAAACTGGT


ACAAAATTCATTACTGAAAATGGTAAGGAAGTCGATACTTTGGAAGCTAAAGATGCTGAAGGTGGTGCTGTT


CTTTCAGGGTTAACAAAAGACAATGGTTTTGTTTTTAACACTGCTAAGTTAAAAGGAATTTACCAAATCGTT


GAATTGAAAGAAAAATCAAACTACGATAACAACGGTTCTATCTTGGCTGATTCAAAAGCAGTTCCAGTTAAA


ATCACTCTGCCATTGGTAAACAACCAAGGTGTTGTTAAAGATGCTCACATTTATCCAAAGAATACTGAAACA


AAACCACAAGTAGATAAGAACTTTGCAGATAAAGATCTTGATTATACTGACAACCGAAAAGACAAAGGTGTT


GTCTCAGCGACAGTTGGTGACAAAAAAGAATACATAGTTGGAACAAAAATTCTTAAAGGCTCAGACTATAAG


AAACTGGTTTGGACTGATAGCATGACTAAAGGTTTGACGTTCAACAACAACGTTAAAGTAACATTGGATGGT


GAAGATTTTCCTGTTTTAAACTACAAACTCGTAACAGATGACCAAGGTTTCCGTCTTGCCTTGAATGCAACA


GGTCTTGCAGCAGTAGCAGCAGCTGCAAAAGACAAAGATGTTGAAATCAAGATCACTTACTCAGCTACGGTG


AACGGCTCCACTACTGTTGAAATTCCAGAAACCAATGATGTTAAATTGGACTATGGTAATAACCCAACGGAA


GAAAGTGAACCACAAGAAGGTACTCCAGCTAACCAAGAAATTAAAGTCATTAAAGACTGGGCAGTAGATGGT


ACAATTACTGATGCTAATGTTGCAGTTAAAGCTATCTTTACCTTGCAAGAAAAACAAACGGATGGTACATGG


GTGAACGTTGCTTCACACGAAGCAACAAAACCATCACGCTTTGAACATACTTTCACAGGTTTGGATAATGCT


AAAACTTACCGCGTTGTCGAACGTGTTAGCGGCTACACTCCAGAATACGTATCATTTAAAAATGGTGTTGTG


ACTATCAAGAACAACAAAAACTCAAATGATCCAACTCCAATCAACCCATCAGAACCAAAAGTGGTGACTTAT


GGACGTAAATTTGTGAAAACAAATCAAGCTAACACTGAACGCTTGGCAGGAGCTACCTTCCTCGTTAAGAAA


GAAGGCAAATACTTGGCACGTAAAGCAGGTGCAGCAACTGCTGAAGCAAAGGCAGCTGTAAAAACTGCTAAA


CTAGCATTGGATGAAGCTGTTAAAGCTTATAACGACTTGACTAAAGAAAAACAAGAAGGCCAAGAAGGTAAA


ACAGCATTGGCTACTGTTGATCAAAAACAAAAAGCTTACAATGACGCTTTTGTTAAAGCTAACTACTCATAT


GAATGGGTTGCAGATAAAAAGGCTGATAATGTTGTTAAATTGATCTCTAACGCCGGTGGTCAATTTGAAATT


ACTGGTTTGGATAAAGGCACTTATGGCTTGGAAGAAACTCAAGCACCAGCAGGTTATGCGACATTGTCAGGT


GATGTAAACTTTGAAGTAACTGCCACATCATATAGCAAAGGGGCTACAACTGACATCGCATATGATAAAGGC


TCTGTAAAAAAAGATGCCCAACAAGTTCAAAACAAAAAAGTAACCATCCCACAAACAGGTGGTATTGGTACA


ATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCAGTAGTTATCATGAAAAAACGTCAATCAGAG


GAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 105):


MKKINKCLTMFSTLLLILTSLFSVAPAFADDATTDTVTLHKIVMPQAAFDNFTEGTKGKNDSDYVGKQINDL


KSYFGSTDAKEIKGAFFVFKNETGTKFITENGKEVDTLEAKDAEGGAVLSGLTKDNGFVFNTAKLKGIYQIV


ELKEKSNYDNNGSILADSKAVPVKITLPLVNNQGVVKDAHIYPKNTETKPQVDKNFADKDLDYTDNRKDKGV


VSATVGDKKEYIVGTKILKGSDYKKLVWTDSMTKGLTFNNNVKVTLDGEDFPVLNYKLVTDDQGFRLALNAT


GLAAVAAAAKDKDVEIKITYSATVNGSTTVEIPETNDVKLDYGNNPTEESEPQEGTPANQEIKVIKDWAVDG


TITDANVAVKAIFTLQEKQTDGTWVNVASHEATKPSRFEHTFTGLDNAKTYRVVERVSGYTPEYVSFKNGVV


TIKNNKNSNDPTPINPSEPKVVTYGRKFVKTNQANTERLAGATFLVKKEGKYLARKAGAATAEAKAAVKTAK


LALDEAVKAYNDLTKEKQEGQEGKTALATVDQKQKAYNDAFVKANYSYEWVADKKADNVVKLISNAGGQFEI


TGLDKGTYGLEETQAPAGYATLSGDVNFEVTATSYSKGATTDIAYDKGSVKKDAQQVQNKKVTIPQTGGIGT


ILFTIIGLSIMLGAVVIMKKRQSEEA


 


Strain IC250


ORF DNA sequence (SEQ ID NO: 148):


ATGAAAAAAATCAACAAATGTCTTACAATGTTCTCGACACTGCTATTGATCTTAACGTCACTATTCTCAGTT


GCACCAGCGTTTGCGGACGACGCAACAACTGATACTGTGACCTTGCACAAGATTGTCATGCCACAAGCTGCA


TTTGATAACTTTACTGAAGGTACAAAAGGTAAGAATGATAGCGATTATGTTGGTAAACAAATTAATGACCTT


AAATCTTATTTTGGCTCAACCGATGCTAAAGAAATCAAGGGTGCTTTCTTTGTTTTCAAAAATGAAACTGGT


ACAAAATTCATTACTGAAAATGGTAAGGAAGTCGATACTTTGGAAGCTAAAGATGCTGAAGGTGGTGCTGTT


CTTTCAGGGTTAACAAAAGACAATGGTTTTGTTTTTAACACTGCTAAGTTAAAAGGAATTTACCAAATCGTT


GAATTGAAAGAAAAATCAAACTACGATAACAACGGTTCTATCTTGGCTGATTCAAAAGCAGTTCCAGTTAAA


ATCACTCTGCCATTGGTAAACAACCAAGGTGTTGTTAAAGATGCTCACATTTATCCAAAGAATACTGAAACA


AAACCACAAGTAGATAAGAACTTTGCAGATAAAGATCTTGATTATACTGACAACCGAAAAGACAAAGGTGTT


GTCTCAGCGACAGTTGGTGACAAAAAAGAATACATAGTTGGAACAAAAATTCTTAAAGGCTCAGACTATAAG


AAACTGGTTTGGACTGATAGCATGACTAAAGGTTTGACGTTCAACAACAACGTTAAAGTAACATTGGATGGT


GAAGATTTTCCTGTTTTAAACTACAAACTCGTAACAGATGACCAAGGTTTCCGTCTTGCCTTGAATGCAACA


GGTCTTGCAGCAGTAGCAGCAGCTGCAAAAGACAAAGATGTTGAAATCAAGATCACTTACTCAGCTACGGTG


AACGGCTCCACTACTGTTGAAATTCCAGAAACCAATGATGTTAAATTGGACTATGGTAATAACCCAACGGAA


GAAAGTGAACCACAAGAAGGTACTCCAGCTAACCAAGAAATTAAAGTCATTAAAGACTGGGCAGTAGATGGT


ACAATTACTGATGCTAATGTTGCAGTTAAAGCTATCTTTACCTTGCAAGAAAAACAAACGGATGGTACATGG


GTGAACGTTGCTTCACACGAAGCAACAAAACCATCACGCTTTGAACATACTTTCACAGGTTTGGATAATGCT


AAAACTTACCGCGTTGTCGAACGTGTTAGCGGCTACACTCCAGAATACGTATCATTTAAAAATGGTGTTGTG


ACTATCAAGAACAACAAAAACTCAAATGATCCAACTCCAATCAACCCATCAGAACCAAAAGTGGTGACTTAT


GGACGTAAATTTGTGAAAACAAATCAAGCTAACACTGAACGCTTGGCAGGAGCTACCTTCCTCGTTAAGAAA


GAAGGCAAATACTTGGCACGTAAAGCAGGTGCAGCAACTGCTGAAGCAAAGGCAGCTGTAAAAACTGCTAAA


CTAGCATTGGATGAAGCTGTTAAAGCTTATAACGACTTGACTAAAGAAAAACAAGAAGGCCAAGAAGGTAAA


ACAGCATTGGCTACTGTTGATCAAAAACAAAAAGCTTACAATGACGCTTTTGTTAAAGCTAACTACTCATAT


GAATGGGTTGCAGATAAAAAGGCTGATAATGTTGTTAAATTGATCTCTAACGCCGGTGGTCAATTTGAAATT


ACTGGTTTGGATAAAGGCACTTATGGCTTGGAAGAAACTCAAGCACCAGCAGGTTATGCGACATTGTCAGGT


GATGTAAACTTTGAAGTAACTGCCACATCATATAGCAAAGGGGCTACAACTGACATCGCATATGATAAAGGC


TCTGTAAAAAAAGATGCCCAACAAGTTCAAAACAAAAAAGTAACCATCCCACAAACAGGTGGTATTGGTACA


ATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCAGTAGTTATCATGAAAAAACGTCAATCAGAG


GAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 106):


MKKINKCLTMFSTLLLILTSLFSVAPAFADDATTDTVTLHKIVMPQAAFDNFTEGTKGKNDSDYVGKQINDL


KSYFGSTDAKEIKGAFFVFKNETGTKFITENGKEVDTLEAKDAEGGAVLSGLTKDNGFVFNTAKLKGIYQIV


ELKEKSNYDNNGSILADSKAVPVKITLPLVNNQGVVKDAHIYPKNTETKPQVDKNFADKDLDYTDNRKDKGV


VSATVGDKKEYIVGTKILKGSDYKKLVWTDSMTKGLTFNNNVKVTLDGEDFPVLNYKLVTDDQGFRLALNAT


GLAAVAAAAKDKDVEIKITYSATVNGSTTVEIPETNDVKLDYGNNPTEESEPQEGTPANQEIKVIKDWAVDG


TITDANVAVKAIFTLQEKQTDGTWVNVASHEATKPSRFEHTFTGLDNAKTYRVVERVSGYTPEYVSFKNGVV


TIKNNKNSNDPTPINPSEPKVVTYGRKFVKTNQANTERLAGATFLVKKEGKYLARKAGAATAEAKAAVKTAK


LALDEAVKAYNDLTKEKQEGQEGKTALATVDQKQKAYNDAFVKANYSYEWVADKKADNVVKLISNAGGQFEI


TGLDKGTYGLEETQAPAGYATLSGDVNFEVTATSYSKGATTDIAYDKGSVKKDAQQVQNKKVTIPQTGGIGT


ILFTIIGLSIMLGAVVIMKKRQSEEA


 


Strain IC251


ORF DNA sequence (SEQ ID NO: 149):


ATGAAAAAAATCAACAAATGTCTTACAATGTTCTCGACACTGCTATTGATCTTAACGTCACTATTCTCAGTT


GCACCAGCGTTTGCGGACGACGCAACAACTGATACTGTGACCTTGCACAAGATTGTCATGCCACAAGCTGCA


TTTGATAACTTTACTGAAGGTACAAAAGGTAAGAATGATAGCGATTATGTTGGTAAACAAATTAATGACCTT


AAATCTTATTTTGGCTCAACCGATGCTAAAGAAATCAAGGGTGCTTTCTTTGTTTTCAAAAATGAAACTGGT


ACAAAATTCATTACTGAAAATGGTAAGGAAGTCGATACTTTGGAAGCTAAAGATGCTGAAGGTGGTGCTGTT


CTTTCAGGGTTAACAAAAGACAATGGTTTTGTTTTTAACACTGCTAAGTTAAAAGGAATTTACCAAATCGTT


GAATTGAAAGAAAAATCAAACTACGATAACAACGGTTCTATCTTGGCTGATTCAAAAGCAGTTCCAGTTAAA


ATCACTCTGCCATTGGTAAACAACCAAGGTGTTGTTAAAGATGCTCACATTTATCCAAAGAATACTGAAACA


AAACCACAAGTAGATAAGAACTTTGCAGATAAAGATCTTGATTATACTGACAACCGAAAAGACAAAGGTGTT


GTCTCAGCGACAGTTGGTGACAAAAAAGAATACATAGTTGGAACAAAAATTCTTAAAGGCTCAGACTATAAG


AAACTGGTTTGGACTGATAGCATGACTAAAGGTTTGACGTTCAACAACAACGTTAAAGTAACATTGGATGGT


GAAGATTTTCCTGTTTTAAACTACAAACTCGTAACAGATGACCAAGGTTTCCGTCTTGCCTTGAATGCAACA


GGTCTTGCAGCAGTAGCAGCAGCTGCAAAAGACAAAGATGTTGAAATCAAGATCACTTACTCAGCTACGGTG


AACGGCTCCACTACTGTTGAAATTCCAGAAACCAATGATGTTAAATTGGACTATGGTAATAACCCAACGGAA


GAAAGTGAACCACAAGAAGGTACTCCAGCTAACCAAGAAATTAAAGTCATTAAAGACTGGGCAGTAGATGGT


ACAATTACTGATGCTAATGTTGCAGTTAAAGCTATCTTTACCTTGCAAGAAAAACAAACGGATGGTACATGG


GTGAACGTTGCTTCACACGAAGCAACAAAACCATCACGCTTTGAACATACTTTCACAGGTTTGGATAATGCT


AAAACTTACCGCGTTGTCGAACGTGTTAGCGGCTACACTCCAGAATACGTATCATTTAAAAATGGTGTTGTG


ACTATCAAGAACAACAAAAACTCAAATGATCCAACTCCAATCAACCCATCAGAACCAAAAGTGGTGACTTAT


GGACGTAAATTTGTGAAAACAAATCAAGCTAACACTGAACGCTTGGCAGGAGCTACCTTCCTCGTTAAGAAA


GAAGGCAAATACTTGGCACGTAAAGCAGGTGCAGCAACTGCTGAAGCAAAGGCAGCTGTAAAAACTGCTAAA


CTAGCATTGGATGAAGCTGTTAAAGCTTATAACGACTTGACTAAAGAAAAACAAGAAGGCCAAGAAGGTAAA


ACAGCATTGGCTACTGTTGATCAAAAACAAAAAGCTTACAATGACGCTTTTGTTAAAGCTAACTACTCATAT


GAATGGGTTGCAGATAAAAAGGCTGATAATGTTGTTAAATTGATCTCTAACGCCGGTGGTCAATTTGAAATT


ACTGGTTTGGATAAAGGCACTTATGGCTTGGAAGAAACTCAAGCACCAGCAGGTTATGCGACATTGTCAGGT


GATGTAAACTTTGAAGTAACTGCCACATCATATAGCAAAGGGGCTACAACTGACATCGCATATGATAAAGGC


TCTGTAAAAAAAGATGCCCAACAAGTTCAAAACAAAAAAGTAACCATCCCACAAACAGGTGGTATTGGTACA


ATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCAGTAGTTATCATGAAAAAACGTCAATCAGAG


GAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 107):


MKKINKCLTMFSTLLLILTSLFSVAPAFADDATTDTVTLHKIVMPQAAFDNFTEGTKGKNDSDYVGKQINDL


KSYFGSTDAKEIKGAFFVFKNETGTKFITENGKEVDTLEAKDAEGGAVLSGLTKDNGFVFNTAKLKGIYQIV


ELKEKSNYDNNGSILADSKAVPVKITLPLVNNQGVVKDAHIYPKNTETKPQVDKNFADKDLDYTDNRKDKGV


VSATVGDKKEYIVGTKILKGSDYKKLVWTDSMTKGLTFNNNVKVTLDGEDFPVLNYKLVTDDQGFRLALNAT


GLAAVAAAAKDKDVEIKITYSATVNGSTTVEIPETNDVKLDYGNNPTEESEPQEGTPANQEIKVIKDWAVDG


TITDANVAVKAIFTLQEKQTDGTWVNVASHEATKPSRFEHTFTGLDNAKTYRVVERVSGYTPEYVSFKNGVV


TIKNNKNSNDPTPINPSEPKVVTYGRKFVKTNQANTERLAGATFLVKKEGKYLARKAGAATAEAKAAVKTAK


LALDEAVKAYNDLTKEKQEGQEGKTALATVDQKQKAYNDAFVKANYSYEWVADKKADNVVKLISNAGGQFEI


TGLDKGTYGLEETQAPAGYATLSGDVNFEVTATSYSKGATTDIAYDKGSVKKDAQQVQNKKVTIPQTGGIGT


ILFTIIGLSIMLGAVVIMKKRQSEEA


 


Strain IC252


ORF DNA sequence (SEQ ID NO: 150):


ATGAAAAAAATCAACAAATATTTTGCAGTCTTCTCGGCATTGCTACTGACCGTAACATCATTGTTCTCAGTT


GCACCAGTGTTTGCGGAAGAAGCAAAAACTACTGACACAGTGACCTTGCACAAGATTGTCATGCCTCGAACT


GCATTTGACGGTTTTACTGCTGGTACAAAGGGTAAGGATAATACTGACTACGTTGGTAAACAAATCGAAGAC


CTTAAAACTTACTTTGGCTCAGGCGAAGCGAAAGAAATCGCAGGTGCTTACTTTGCTTTCAAAAATGAAGCT


GGTACTAAATACATCACTGAAAATGGTGAAGAAGTTGATACTTTGGATACAACAGATGCCAAAGGTGGTGCT


GTTCTTAAAGGTTTAACAACAGACAATGGTTTCAAATTTAACACTTCTAAATTAACAGGAACTTACCAAATC


GTTGAATTGAAAGAAAAATCTACATACAACAACGATGGTTCTATCTTGGCTGATTCAAAAGCAGTTCCAGTT


AAAATCACTCTTCCATTGGTAAACGACAATGGTGTTGTTAAAGACGCTCACGTTTATCCAAAGAACACTGAA


ACAAAACCACAAGTAGATAAGAACTTCGCAGATAAAGAACTTGATTATGCGAACAACAAAAAAGACAAAGGG


ACTGTCTCAGCATCTGTTGGTGATGTTAAAAAATATCATGTTGGAACAAAAATCCTTAAAGGTTCAGACTAT


AAGAAATTAATCTGGACCGATAGCATGACCAAAGGTTTGACTTTCAACAACGATATTGCTGTAACATTGGAT


GGTGCAACTCTTGATGCTACAAATTACAAACTTGTAGCAGATGACCAAGGTTTCCGCCTTGTCTTGACTGAC


AAAGGTCTTGAAGCAGTGGCAAAAGCCGCAAAAACAAAAGATGTTGAAATCAAGATCACTTACTCAGCTACT


TTGAACGGTTCTGCTGTCGTTGAAGTTCTAGAAACCAATGATGTTAAATTGGACTACGGCAACAACCCAACA


ATTGAAAATGAACCAAAAGAAGGTATTCCAGTTGATAAGAAAATCACTGTTAACAAAACATGGGCAGTAGAT


GGCAATGAAGTGAATAAAGCAGATGAAACAGTTGATGCTGTCTTCACCTTGCAAGTTAAAGATGGTGACAAA


TGGGTGAATGTTGATTCAGCTAAAGCAACAGCTGCAACTAGCTTCAAACACACTTTTGAAAACTTGGATAAT


GCTAAAACTTACCGCGTTATCGAACGTGTTAGCGGCTACGCTCCAGAATACGTCTCATTTGTAAATGGCGTT


GTAACCATCAAGAACAACAAAGACTCAAATGAGCCAACTCCAATCAACCCATCAGAACCAAAAGTGGTGACT


TATGGACGTAAATTTGTGAAAACAAATAAAGATGGAAAAGAACGCTTGGCAGGAGCTACCTTCCTTGTTAAG


AAAGATGGCAAGTACTTGGCACGTAAATCAGGTGTTGCAACAGATGCAGAAAAAGCTGCTGTAGATTCAACT


AAATCAGCATTGGATGCTGCTGTTAAAGCTTACAATGATTTGACTAAAGAAAAACAAGAAGGTCAAGATGGT


AAATCAGCATTGGCTACCGTTAGTGAAAAACAAAAAGCTTACAATGATGCCTTTGTTAAAGCTAACTACTCA


TACGAATGGGTTGAAGATAAAAATGCTAAGAATGTTGTTAAATTGATTTCTAACGATAAAGGTCAATTTGAA


ATTACTGGCTTGACTGAAGGTCAATACTCATTGGAAGAAACACAAGCACCAACTGGTTATGCTAAATTATCA


GGTGATGTTTCGTTTAATGTTAATGCTACTTCATACAGTAAAGGTTCTGCTCAAGATATTGAGTATACCCAA


GGTTCTAAAACTAAAGATGCACAACAAGTTATCAATAAGAAGGTTACTATTCCACAAACAGGTGGTATTGGT


ACAATTTTTTTCACAATTATTGGATTAAGTATTATGCTTGGAGCGGTAGTTATCATGAAAAGACGTCAATCA


GAGGAAGTTTAA


 


ORF amino acid sequence (SEQ ID NO: 108):


MKKINKCLTMFSTLLLILTSLFSVAPAFADDATTDTVTLHKIVMPQAAFDNFTEGTKGKNDSDYVGKQINDL


KSYFGSTDAKEIKGAFFVFKNETGTKFITENGKEVDTLEAKDAEGGAVLSGLTKDNGFVFNTAKLKGIYQIV


ELKEKSNYDNNGSILADSKAVPVKITLPLVNNQGVVKDAHIYPKNTETKPQVDKNFADKDLDYTDNRKDKGV


VSATVGDKKEYIVGTKILKGSDYKKLVWTDSMTKGLTFNNNVKVTLDGEDFPVLNYKLVTDDQGFRLALNAT


GLAAVAAAAKDKDVEIKITYSATVNGSTTVEIPETNDVKLDYGNNPTEESEPQEGTPANQEIKVIKDWAVDG


TITDANVAVKAIFTLQEKQTDGTWVNVASHEATKPSRFEHTFTGLDNAKTYRVVERVSGYTPEYVSFKNGVV


TIKNNKNSNDPTPINPSEPKVVTYGRKFVKTNQANTERLAGATFLVKKEGKYLARKAGAATAEAKAAVKTAK


LALDEAVKAYNDLTKEKQEGQEGKTALATVDQKQKAYNDAFVKANYSYEWVADKKADNVVKLISNAGGQFEI


TGLDKGTYGLEETQAPAGYATLSGDVNFEVTATSYSKGATTDIAYDKGSVKKDAQQVQNKKVTIPQTGGIGT


ILFTIIGLSIMLGAVVIMKKRQSEEA


 


Strain IC253


ORF DNA sequence (SEQ ID NO: 151):


ATGAAAAAAATCAACAAATGTCTTACAGTGTTCTCGACACTGCTATTGATCTTAACGTCACTATTCTCAGTT


GCACCAGCGTTTGCGGACGACGTAACAACTGATACTGTGACCTTGCACAAGATTGTCATGCCACAAGCTGCA


TTTGATAACTTTACTGAAGGTACAAAAGGTAAGAATGATAGCGATTATGTTGGTAAACAAATTAATGACCTT


AAATCTTATTTTGGCTCAACCGATGCTAAAGAAATTAAGGGTGCTTTCTTTGTTTTCAAAAATGAAACTGGT


ACAAAATTCATTACTGAAAATGGTAAGGAAGTCGATACTTTGGAAGCTAAAGATGCTGAAGGTGGTGCTGTT


CTTTCAGGGTTAACAAAAGACACTGGTTTTGCTTTTAACACTGCTAAGTTAAAAGGAACTTACCAAATCGTT


GAATTGAAAGAAAAATCAAACTACGATAACAACGGTTCTATCTTGGCTGATTCAAAAGCAGTTCCAGTTAAA


ATCACTCTGCCATTGGTAAACAACCAAGGTGTTGTTAAAGATGCTCACATTTATCCAAAGAATACTGAAACA


AAACCACAAGTAGATAAGAACTTTGCAGATAAAGATCTTGATTATACTGACAACCGAAAAGACAAAGGTGTT


GTCTCAGCGACAGTTGGTGACAAAAAAGAATACATAGTTGGAACAAAAATTCTTAAAGGCTCAGACTATAAG


AAACTGGTTTGGACTGATAGCATGACTAAAGGTTTGACGTTCAACAACAACGTTAAAGTAACATTGGATGGT


AAAGATTTTCCTGTTTTAAACTACAAACTCGTAACAGATGACCAAGGTTTCCGTCTTGCCTTGAATGCAACA


GGTCTTGCAGCAGTAGCAGCTGCTGCAAAAGACAAAGATGTTGAAATCAAGATCACTTACTCAGCTACGGTG


AACGGCTCCACTACTGTTGAAGTTCCAGAAACCAATGATGTTAAATTGGACTATGGTAATAACCCAACGGAA


GAAAGTGAACCACAAGAAGGTACTCCAGCTAACCAAGAAATTAAAGTCATTAAAGACTGGGCAGTAGATGGT


ACAATTACTGATGTTAATGTTGCAGTTAAAGCTATCTTTACCTTGCAAGAAAAACAAACGGATGGTACATGG


GTGAACGTTGCTTCACACGAAGCAACAAAACCATCACGCTTTGAACATACTTTCACAGGTTTGGATAATACT


AAAACTTACCGCGTTGTCGAACGTGTTAGCGGCTACACTCCAGAATATGTATCATTTAAAAATGGTGTTGTG


ACTATCAAGAACAACAAAAACTCAAATGATCCAACTCCAATCAACCCATCAGAACCAAAAGTGGTGACTTAT


GGACGTAAATTTGTGAAAACAAATCAAGCTAACACTGAACGCTTGGCAGGAGCTACCTTCCTTGTTAAGAAA


GAAGGAAAATACTTGGCACGTAAAGCAGGTGCAGCAACTGCTGAAGCAAAGGCAGCTGTAAAAACTGCTAAA


CTAGCATTGGATGAAGCTGTTAAAGCTTATAACGACTTGACTAAAGAAAAACAAGAAGGCCAAGAAGGTAAA


ACAGCATTGGCTACTGTTGATCAAAAACAAAAAGCTTACAATGACGCTTTTGTTAAAGCTAACTACTCATAT


GAATGGGTTGCAGATAAAAAGGCTGATAATGTTGTTAAATTGATCTCTAACGCCGGTGGTCAATTTGAAATT


ACTGGTTTGGATAAAGGCACTTATAGCTTGGAAGAAACTCAAGCACCAGCAGGTTATGCGACATTGTCAGGT


GATGTAAACTTTGAAGTAACTGCCACATCATATAGCAAAGGGGCTACAACTGACATCGCATATGATAAAGGA


TCTGTAAAAAAAGATGCCCAACAAGTTCAAAACAAAAAAGTAACCATCCCACAAACAGGTGGTATTGGTACA


ATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCAGTAGTTGTCATGAAAAAACGTCAATCAGAG


GAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 109):


MKKINKCLTVFSTLLLILTSLFSVAPAFADDVTTDTVTLHKIVMPQAAFDNFTEGTKGKNDSDYVGKQINDL


KSYFGSTDAKEIKGAFFVFKNETGTKFITENGKEVDTLEAKDAEGGAVLSGLTKDTGFAFNTAKLKGTYQIV


ELKEKSNYDNNGSILADSKAVPVKITLPLVNNQGVVKDAHIYPKNTETKPQVDKNFADKDLDYTDNRKDKGV


VSATVGDKKEYIVGTKILKGSDYKKLVWTDSMTKGLTFNNNVKVTLDGKDFPVLNYKLVTDDQGFRLALNAT


GLAAVAAAAKDKDVEIKITYSATVNGSTTVEVPETNDVKLDYGNNPTEESEPQEGTPANQEIKVIKDWAVDG


TITDVNVAVKAIFTLQEKQTDGTWVNVASHEATKPSRFEHTFTGLDNTKTYRVVERVSGYTPEYVSFKNGVV


TIKNNKNSNDPTPINPSEPKVVTYGRKFVKTNQANTERLAGATFLVKKEGKYLARKAGAATAEAKAAVKTAK


LALDEAVKAYNDLTKEKQEGQEGKTALATVDQKQKAYNDAFVKANYSYEWVADKKADNVVKLISNAGGQFEI


TGLDKGTYSLEETQAPAGYATLSGDVNFEVTATSYSKGATTDIAYDKGSVKKDAQQVQNKKVTIPQTGGIGT


ILFTIIGLSIMLGAVVVMKKRQSEEA


 


Strain IC254


ORF DNA sequence (SEQ ID NO: 152):


ATGAAAAGAATCAACAAATATTTTGCAATGTTCTCGGCATTGTTACTGACTTTAACGTCATTGCTCTCAGTT


GCACCAGCGTTTGCGGATGAAGCAACAACTAATACAGTGACTTTGCACAAGATTTTGCAAACCGAATCAAAT


CTTAACAAAAGTAACTTCCCAGGAACTACAGGTCTTAACGGAAAAGACTACAAAGGTGGAGCTATTTCTGAC


CTTGCTGGTTACTTTGGCGAGGGATCTAAAGAAATCGAAGGTGCGTTCTTTGCTTTAGCTTTGAAAGAAGAT


AAAAGTGGTAAAGTGCAATATGTTAAGGCAAAAGAAGGTAACAAATTAACACCAGCCTTAATTAATAAAGAT


GGTACTCCTGAAATAACAGTAAATATTGATGAGGCCGTGTCTGGATTGACACCAGAGGGAGATACTGGACTT


GTTTTCAACACCAAAGGATTGAAAGGCGAGTTTAAAATTGTTGAAGTTAAATCAAAATCTACTTACAACAAT


AATGGTTCCCTCCTGGCTGCTTCAAAAGCGGTTCCAGTTAACATCACTCTTCCATTGGTAAATGAAGATGGT


GTTGTTGCTGATGCCCATGTTTATCCAAAGAACACTGAAGAAAAACCAGAAATTGATAAAAACTTTGCTAAA


ACAAACGATTTGACAGCATTGACAGATGTTAATAGACTTTTGACAGCTGGCGCAAATTATGGTAATTATGCA


CGTGACAAAGCAACTGCTACTGCTGAAATCGGTAAAGTTGTTCCTTATGAAGTTAAAACAAAAATTCACAAA


GGTTCTAAATACGAAAACTTGGTTTGGACAGATATAATGTCAAATGGTTTGACAATGGGTTCAACTGTTAGC


CTTAAAGCTTCAGGAACTACAGAAACTTTTGCTAAGGATACAGACTATGAACTTAGCATTGATGCCCGTGGT


TTCACATTAAAATTCACAGCTGATGGATTGGGCAAATTGGAAAAAGCAGCTAAAACAGCTGATATTGAATTT


ACATTGACTTATAGTGCTACTGTTAATGGTCAAGCAATTATTGATAATCCAGAATCCAATGATATCAAATTG


TCGTATGGTAACAAACCAGGTAAAGACTTGACTGAACTTCCTGTTACACCTTCAAAGGGTGAAGTAACAGTT


GCTAAAACTTGGTCTGACGGAATTGCACCTGATGGTGTAAACGTTGTTTACACATTGAAAGATAAAGATAAA


ACTGTTGCTTCAGTATCATTGACAAAAACATCTAAAGGTACAATCGACCTTGGAAATGGTATCAAATTTGAA


GTATCTGGTAACTTCTCGGGTAAATTCACTGGTCTAGAAAACAAATCATACATGATCTCAGAACGTGTTTCT


GGTTACGGAAGTGCAATAAATCTAGAAAATGGTAAAGTAACCATTACCAATACCAAAGATTCTGATAACCCA


ACACCATTGAACCCAACTGAACCAAAAGTTGAAACTCATGGTAAGAAATTTGTCAAAACTAATGAACAAGGT


GACCGTTTGGCTGGTGCACAATTCGTTGTGAAAAACTCAGCAGGTAAATACCTTGCTCTTAAAGCAGATCAA


TCAGAAGGTCAAAAAACTTTAGCTGCTAAGAAAATAGCTTTAGATGAAGCTATCGCTGCTTATAACAAGTTG


TCTGCAACAGACCAAAAAGGTGAAAAAGGAATTACTGCAAAAGAACTTATCAAAACTAAACAAGCAGATTAC


GATGCAGCCTTCATTGAGGCTCGTACAGCTTATGAGTGGATAACAGATAAGGCTAGAGCCATTACCTACACT


TCAAACGATCAAGGTCAATTTGAAGTTACAGGTCTTGCAGACGGTACTTACAACCTTGAAGAAACACTTGCT


CCAGCAGGATTTGCTAAGTTGGCAGGTAATATTAAGTTTGTAGTTAATCAAGGGTCATACATAACAGGTGGT


AACATTGACTACGTTGCTAACAGCAACCAAAAAGATGCGACACGTGTAGAAAATAAAAAGGTAACAATCCCA


CAAACAGGTGGTATTGGTACAATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCAGTAGTTATC


ATGAAAAGACGCCAATCAAAGGAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 110):


MKRINKYFAMFSALLLTLTSLLSVAPAFADEATTNTVTLHKILQTESNLNKSNFPGTTGLNGKDYKGGAISD


LAGYFGEGSKEIEGAFFALALKEDKSGKVQYVKAKEGNKLTPALINKDGTPEITVNIDEAVSGLTPEGDTGL


VFNTKGLKGEFKIVEVKSKSTYNNNGSLLAASKAVPVNITLPLVNEDGVVADAHVYPKNTEEKPEIDKNFAK


TNDLTALTDVNRLLTAGANYGNYARDKATATAEIGKVVPYEVKTKIHKGSKYENLVWTDIMSNGLTMGSTVS


LKASGTTETFAKDTDYELSIDARGFTLKFTADGLGKLEKAAKTADIEFTLTYSATVNGQAIIDNPESNDIKL


SYGNKPGKDLTELPVTPSKGEVTVAKTWSDGIAPDGVNVVYTLKDKDKTVASVSLTKTSKGTIDLGNGIKFE


VSGNFSGKFTGLENKSYMISERVSGYGSAINLENGKVTITNTKDSDNPTPLNPTEPKVETHGKKFVKTNEQG


DRLAGAQFVVKNSAGKYLALKADQSEGQKTLAAKKIALDEAIAAYNKLSATDQKGEKGITAKELIKTKQADY


DAAFIEARTAYEWITDKARAITYTSNDQGQFEVTGLADGTYNLEETLAPAGFAKLAGNIKFVVNQGSYITGG


NIDYVANSNQKDATRVENKKVTIPQTGGIGTILFTIIGLSIMLGAVVIMKRRQSKEA


 


Strain IC255


ORF DNA sequence (SEQ ID NO: 153):


ATGAAAAAAATCAACAAATGTCTTACAATGTTCTCGACACTGCTATTGATCTTAACGTCACTATTCTCAGTT


GCACCAGCGTTTGCGGACGACGCAACAACTGATACTGTGACCTTGCACAAGATTGTCATGCCACAAGCTGCA


TTTGATAACTTTACTGAAGGTACAAAAGGTAAGAATGATAGCGATTATGTTGGTAAACAAATTAATGACCTT


AAATCTTATTTTGGCTCAACCGATGCTAAAGAAATCAAGGGTGCTTTCTTTGTTTTCAAAAATGAAACTGGT


ACAAAATTCATTACTGAAAATGGTAAGGAAGTCGATACTTTGGAAGCTAAAGATGCTGAAGGTGGTGCTGTT


CTTTCAGGGTTAACAAAAGACAATGGTTTTGTTTTTAACACTGCTAAGTTAAAAGGAATTTACCAAATCGTT


GAATTGAAAGAAAAATCAAACTACGATAACAACGGTTCTATCTTGGCTGATTCAAAAGCAGTTCCAGTTAAA


ATCACTCTGCCATTGGTAAACAACCAAGGTGTTGTTAAAGATGCTCACATTTATCCAAAGAATACTGAAACA


AAACCACAAGTAGATAAGAACTTTGCAGATAAAGATCTTGATTATACTGACAACCGAAAAGACAAAGGTGTT


GTCTCAGCGACAGTTGGTGACAAAAAAGAATACATAGTTGGAACAAAAATTCTTAAAGGCTCAGACTATAAG


AAACTGGTTTGGACTGATAGCATGACTAAAGGTTTGACGTTCAACAACAACGTTAAAGTAACATTGGATGGT


GAAGATTTTCCTGTTTTAAACTACAAACTCGTAACAGATGACCAAGGTTTCCGTCTTGCCTTGAATGCAACA


GGTCTTGCAGCAGTAGCAGCAGCTGCAAAAGACAAAGATGTTGAAATCAAGATCACTTACTCAGCTACGGTG


AACGGCTCCACTACTGTTGAAATTCCAGAAACCAATGATGTTAAATTGGACTATGGTAATAACCCAACGGAA


GAAAGTGAACCACAAGAAGGTACTCCAGCTAACCAAGAAATTAAAGTCATTAAAGACTGGGCAGTAGATGGT


ACAATTACTGATGCTAATGTTGCAGTTAAAGCTATCTTTACCTTGCAAGAAAAACAAACGGATGGTACATGG


GTGAACGTTGCTTCACACGAAGCAACAAAACCATCACGCTTTGAACATACTTTCACAGGTTTGGATAATGCT


AAAACTTACCGCGTTGTCGAACGTGTTAGCGGCTACACTCCAGAATACGTATCATTTAAAAATGGTGTTGTG


ACTATCAAGAACAACAAAAACTCAAATGATCCAACTCCAATCAACCCATCAGAACCAAAAGTGGTGACTTAT


GGACGTAAATTTGTGAAAACAAATCAAGCTAACACTGAACGCTTGGCAGGAGCTACCTTCCTCGTTAAGAAA


GAAGGCAAATACTTGGCACGTAAAGCAGGTGCAGCAACTGCTGAAGCAAAGGCAGCTGTAAAAACTGCTAAA


CTAGCATTGGATGAAGCTGTTAAAGCTTATAACGACTTGACTAAAGAAAAACAAGAAGGCCAAGAAGGTAAA


ACAGCATTGGCTACTGTTGATCAAAAACAAAAAGCTTACAATGACGCTTTTGTTAAAGCTAACTACTCATAT


GAATGGGTTGCAGATAAAAAGGCTGATAATGTTGTTAAATTGATCTCTAACGCCGGTGGTCAATTTGAAATT


ACTGGTTTGGATAAAGGCACTTATGGCTTGGAAGAAACTCAAGCACCAGCAGGTTATGCGACATTGTCAGGT


GATGTAAACTTTGAAGTAACTGCCACATCATATAGCAAAGGGGCTACAACTGACATCGCATATGATAAAGGC


TCTGTAAAAAAAGATGCCCAACAAGTTCAAAACAAAAAAGTAACCATCCCACAAACAGGTGGTATTGGTACA


ATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCAGTAGTTATCATGAAAAAACGTCAATCAGAG


GAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 111):


MKKINKCLTMFSTLLLILTSLFSVAPAFADDATTDTVTLHKIVMPQAAFDNFTEGTKGKNDSDYVGKQINDL


KSYFGSTDAKEIKGAFFVFKNETGTKFITENGKEVDTLEAKDAEGGAVLSGLTKDNGFVFNTAKLKGIYQIV


ELKEKSNYDNNGSILADSKAVPVKITLPLVNNQGVVKDAHIYPKNTETKPQVDKNFADKDLDYTDNRKDKGV


VSATVGDKKEYIVGTKILKGSDYKKLVWTDSMTKGLTFNNNVKVTLDGEDFPVLNYKLVTDDQGFRLALNAT


GLAAVAAAAKDKDVEIKITYSATVNGSTTVEIPETNDVKLDYGNNPTEESEPQEGTPANQEIKVIKDWAVDG


TITDANVAVKAIFTLQEKQTDGTWVNVASHEATKPSRFEHTFTGLDNAKTYRVVERVSGYTPEYVSFKNGVV


TIKNNKNSNDPTPINPSEPKVVTYGRKFVKTNQANTERLAGATFLVKKEGKYLARKAGAATAEAKAAVKTAK


LALDEAVKAYNDLTKEKQEGQEGKTALATVDQKQKAYNDAFVKANYSYEWVADKKADNVVKLISNAGGQFEI


TGLDKGTYGLEETQAPAGYATLSGDVNFEVTATSYSKGATTDIAYDKGSVKKDAQQVQNKKVTIPQTGGIGT


ILFTIIGLSIMLGAVVIMKKRQSEEA


 


Strain IC287


ORF DNA sequence (SEQ ID NO: 154):


ATGAAAAAAATCAACAAATGTCTTACAATGTTCTCGACACTGCTATTGATCTTAACGTCACTATTCTCAGTT


GCACCAGCGTTTGCGGACGACGCAACAACTGATACTGTGACCTTGCACAAGATTGTCATGCCACAAGCTGCA


TTTGATAACTTTACTGAAGGTACAAAAGGTAAGAATGATAGCGATTATGTTGGTAAACAAATTAATGACCTT


AAATCTTATTTTGGCTCAACCGATGCTAAAGAAATCAAGGGTGCTTTCTTTGTTTTCAAAAATGAAACTGGT


ACAAAATTCATTACTGAAAATGGTAAGGAAGTCGATACTTTGGAAGCTAAAGATGCTGAAGGTGGTGCTGTT


CTTTCAGGGTTAACAAAAGACAATGGTTTTGTTTTTAACACTGCTAAGTTAAAAGGAATTTACCAAATCGTT


GAATTGAAAGAAAAATCAAACTACGATAACAACGGTTCTATCTTGGCTGATTCAAAAGCAGTTCCAGTTAAA


ATCACTCTGCCATTGGTAAACAACCAAGGTGTTGTTAAAGATGCTCACATTTATCCAAAGAATACTGAAACA


AAACCACAAGTAGATAAGAACTTTGCAGATAAAGATCTTGATTATACTGACAACCGAAAAGACAAAGGTGTT


GTCTCAGCGACAGTTGGTGACAAAAAAGAATACATAGTTGGAACAAAAATTCTTAAAGGCTCAGACTATAAG


AAACTGGTTTGGACTGATAGCATGACTAAAGGTTTGACGTTCAACAACAACGTTAAAGTAACATTGGATGGT


GAAGATTTTCCTGTTTTAAACTACAAACTCGTAACAGATGACCAAGGTTTCCGTCTTGCCTTGAATGCAACA


GGTCTTGCAGCAGTAGCAGCAGCTGCAAAAGACAAAGATGTTGAAATCAAGATCACTTACTCAGCTACGGTG


AACGGCTCCACTACTGTTGAAATTCCAGAAACCAATGATGTTAAATTGGACTATGGTAATAACCCAACGGAA


GAAAGTGAACCACAAGAAGGTACTCCAGCTAACCAAGAAATTAAAGTCATTAAAGACTGGGCAGTAGATGGT


ACAATTACTGATGCTAATGTTGCAGTTAAAGCTATCTTTACCTTGCAAGAAAAACAAACGGATGGTACATGG


GTGAACGTTGCTTCACACGAAGCAACAAAACCATCACGCTTTGAACATACTTTCACAGGTTTGGATAATGCT


AAAACTTACCGCGTTGTCGAACGTGTTAGCGGCTACACTCCAGAATACGTATCATTTAAAAATGGTGTTGTG


ACTATCAAGAACAACAAAAACTCAAATGATCCAACTCCAATCAACCCATCAGAACCAAAAGTGGTGACTTAT


GGACGTAAATTTGTGAAAACAAATCAAGCTAACACTGAACGCTTGGCAGGAGCTACCTTCCTCGTTAAGAAA


GAAGGCAAATACTTGGCACGTAAAGCAGGTGCAGCAACTGCTGAAGCAAAGGCAGCTGTAAAAACTGCTAAA


CTAGCATTGGATGAAGCTGTTAAAGCTTATAACGACTTGACTAAAGAAAAACAAGAAGGCCAAGAAGGTAAA


ACAGCATTGGCTACTGTTGATCAAAAACAAAAAGCTTACAATGACGCTTTTGTTAAAGCTAACTACTCATAT


GAATGGGTTGCAGATAAAAAGGCTGATAATGTTGTTAAATTGATCTCTAACGCCGGTGGTCAATTTGAAATT


ACTGGTTTGGATAAAGGCACTTATGGCTTGGAAGAAACTCAAGCACCAGCAGGTTATGCGACATTGTCAGGT


GATGTAAACTTTGAAGTAACTGCCACATCATATAGCAAAGGGGCTACAACTGACATCGCATATGATAAAGGC


TCTGTAAAAAAAGATGCCCAACAAGTTCAAAACAAAAAAGTAACCATCCCACAAACAGGTGGTATTGGTACA


ATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCAGTAGTTATCATGAAAAAACGTCAATCAGAG


GAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 112):


MKKINKCLTMFSTLLLILTSLFSVAPAFADDATTDTVTLHKIVMPQAAFDNFTEGTKGKNDSDYVGKQINDL


KSYFGSTDAKEIKGAFFVFKNETGTKFITENGKEVDTLEAKDAEGGAVLSGLTKDNGFVFNTAKLKGIYQIV


ELKEKSNYDNNGSILADSKAVPVKITLPLVNNQGVVKDAHIYPKNTETKPQVDKNFADKDLDYTDNRKDKGV


VSATVGDKKEYIVGTKILKGSDYKKLVWTDSMTKGLTFNNNVKVTLDGEDFPVLNYKLVTDDQGFRLALNAT


GLAAVAAAAKDKDVEIKITYSATVNGSTTVEIPETNDVKLDYGNNPTEESEPQEGTPANQEIKVIKDWAVDG


TITDANVAVKAIFTLQEKQTDGTWVNVASHEATKPSRFEHTFTGLDNAKTYRVVERVSGYTPEYVSFKNGVV


TIKNNKNSNDPTPINPSEPKVVTYGRKFVKTNQANTERLAGATFLVKKEGKYLARKAGAATAEAKAAVKTAK


LALDEAVKAYNDLTKEKQEGQEGKTALATVDQKQKAYNDAFVKANYSYEWVADKKADNVVKLISNAGGQFEI


TGLDKGTYGLEETQAPAGYATLSGDVNFEVTATSYSKGATTDIAYDKGSVKKDAQQVQNKKVTIPQTGGIGT


ILFTIIGLSIMLGAVVIMKKRQSEEA


 


Strain IC289


ORF DNA sequence (SEQ ID NO: 155):


ATGAAAAGAATCAACAAATATTTTGCAATGTTCTCGGCATTGTTATTAATTTTAACATCGTTGTTATCGGTA


GCTCCGGTATTTGCTGCTGAGATGGGAAATATCACTAAAACAGTAACCTTACACAAAATTGTTCAAACATCC


GATAATTTGGCTAAGCCAAATTTCCCAGGAATAAATGGATTGAATGGAACGAAGTATATGGGTCAAAAACTT


ACTGACATTTCAGGATATTTTGGGCAAGGTTCTAAAGAAATCGCCGGTGCTTTCTTTGCGGTTATGAATGAA


AGTCAGACAAAATATATCACAGAAAGTGGTACTGAAGTAGAAAGTATCGATGCAGCAGGTGTCCTTAAAGGT


TTGACAACTGAAAACGGCATTACATTTAATACTGCAAACTTAAAAGGAACATACCAAATCGTTGAGTTGCTT


GACAAATCTAATTATAAAAATGGTGACAAAGTTCTTGCTGACTCAAAAGCTGTCCCAGTGAAAATCACTCTT


CCTTTGTATAACGAAGAAGGAATTGTCGTGGACGCTGAAGTGTATCCAAAGAATACAGAAGAAGCACCACAA


ATCGACAAAAACTTTGCTAAAGCAAATAAATTGTTGAATGACAGTGATAATTCAGCTATTGCAGGTGGGGCA


GACTACGACAAATATCAGGCAGAAAAAGCAAAAGCTACTGCTGAAATCGGTCAAGAAATCCCTTACGAAGTT


AAAACAAAAATCCAAAAAGGGTCTAAATACAAAAACCTTGCTTGGGTCGATACCATGTCAAATGGTTTGACA


ATGGGTAACACTGTTAACTTAGAAGCATCGTCAGGCTCTTTTGTAGAAGGTACAGATTACAATGTTGAACGT


GATGACCGTGGTTTCACTTTGAAATTCACAGATACAGGTTTGACTAAGCTACAAAAAGAAGCGGAAACACAA


GCTGTTGAATTCACATTGACATATAGCGCAACAGTTAACGGTGCGGCTATTGATGACAAGCCAGAAAGCAAT


GATATCAAACTTCAATACGGTAACAAACCAGGTAAAAAAGTAAAAGAAATCCCAGTAACACCGTCAAATGGC


GAAATCACTGTTAGCAAAACTTGGGACAAAGGTTCAGATTTAGAGAATGCGAATGTTGTTTATACCCTTAAA


GATGGTGGAACAGCTGTTGCCTCAGTTTCATTGACAAAAACAACACCAAATGGCGAAATCAACTTAGGTAAT


GGTATTAAATTTACAGTTACTGGAGCGTTTGCTGGTAAATTCAGTGGTCTGACTGATAGTAAAACATACATG


ATCTCAGAACGTATCGCTGGTTATGGTAATACAATCACTACTGGTGCTGGTAGTGCAGCTATCACCAATACT


CCAGATTCAGACAACCCAACACCACTTAATCCAACTGAACCAAAAGTTGTGACACACGGTAAAAAATTCGTC


AAAACAAGTTCGACTGAAACAGAACGCTTGCAAGGTGCACAGTTCGTTGTTAAAGATTCAGCTGGTAAATAC


CTTGCATTGAAATCATCTGCGACAATATCAGCTCAAACAACAGCTTACACAAATGCTAAAACTGCTCTTGAC


GCTAAAATCGCAGCTTACAACAAACTTTCAGCAGACGATCAAAAAGGTACTAAAGGTGAAACAGCTAAAGCA


GAAATCAAAACTGCTCAAGACGCTTACAATGCAGCCTTCATCGTAGCTCGTACAGCTTACGAGTGGGTAACT


AATAAAGAAGATGCTAACGTTGTTAAAGTGACTTCAAACGCTGACGGTCAATTTGAAGTTAGCGGTCTTGCA


ACTGGTGATTATAAACTTGAAGAAACACAAGCTCCAGCTGGTTACGCTAAATTAGCAGGTGATGTTGATTTC


AAAGTTGGAAACAGCTCAAAAGCAGACGACTCAGGTAACATTGATTACACTGCTAGCAGCAATAAAAAAGAC


GCTCAACGCATAGAAAACAAAAAAGTGACTATTCCACAAACAGGTGGTATTGGTACAATTCTTTTCACAATT


ATTGGTTTAAGCATTATGCTTGGAGCGGTAATTATCATGAAAAGACGTCAATCAGAGGAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 113):


MKRINKYFAMFSALLLILTSLLSVAPVFAAEMGNITKTVTLHKIVQTSDNLAKPNFPGINGLNGTKYMGQKL


TDISGYFGQGSKEIAGAFFAVMNESQTKYITESGTEVESIDAAGVLKGLTTENGITFNTANLKGTYQIVELL


DKSNYKNGDKVLADSKAVPVKITLPLYNEEGIVVDAEVYPKNTEEAPQIDKNFAKANKLLNDSDNSAIAGGA


DYDKYQAEKAKATAEIGQEIPYEVKTKIQKGSKYKNLAWVDTMSNGLTMGNTVNLEASSGSFVEGTDYNVER


DDRGFTLKFTDTGLTKLQKEAETQAVEFTLTYSATVNGAAIDDKPESNDIKLQYGNKPGKKVKEIPVTPSNG


EITVSKTWDKGSDLENANVVYTLKDGGTAVASVSLTKTTPNGEINLGNGIKFTVTGAFAGKFSGLTDSKTYM


ISERIAGYGNTITTGAGSAAITNTPDSDNPTPLNPTEPKVVTHGKKFVKTSSTETERLQGAQFVVKDSAGKY


LALKSSATISAQTTAYTNAKTALDAKIAAYNKLSADDQKGTKGETAKAEIKTAQDAYNAAFIVARTAYEWVT


NKEDANVVKVTSNADGQFEVSGLATGDYKLEETQAPAGYAKLAGDVDFKVGNSSKADDSGNIDYTASSNKKD


AQRIENKKVTIPQTGGIGTILFTIIGLSIMLGAVIIMKRRQSEEA


 


Strain IC291


ORF DNA sequence (SEQ ID NO: 156):


ATGAAAAAAATCAACAAATGTCTTACAATGTTCTCGACACTGCTATTGATCTTAACGTCACTATTCTCAGTT


GCACCAGCGTTTGCGGACGACGCAACAACTGATACTGTGACCTTGCACAAGATTGTCATGCCACAAGCTGCA


TTTGATAACTTTACTGAAGGTACAAAAGGTAAGAATGATAGCGATTATGTTGGTAAACAAATTAATGACCTT


AAATCTTATTTTGGCTCAACCGATGCTAAAGAAATCAAGGGTGCTTTCTTTGTTTTCAAAAATGAAACTGGT


ACAAAATTCATTACTGAAAATGGTAAGGAAGTCGATACTTTGGAAGCTAAAGATGCTGAAGGTGGTGCTGTT


CTTTCAGGGTTAACAAAAGACAATGGTTTTGTTTTTAACACTGCTAAGTTAAAAGGAATTTACCAAATCGTT


GAATTGAAAGAAAAATCAAACTACGATAACAACGGTTCTATCTTGGCTGATTCAAAAGCAGTTCCAGTTAAA


ATCACTCTGCCATTGGTAAACAACCAAGGTGTTGTTAAAGATGCTCACATTTATCCAAAGAATACTGAAACA


AAACCACAAGTAGATAAGAACTTTGCAGATAAAGATCTTGATTATACTGACAACCGAAAAGACAAAGGTGTT


GTCTCAGCGACAGTTGGTGACAAAAAAGAATACATAGTTGGAACAAAAATTCTTAAAGGCTCAGACTATAAG


AAACTGGTTTGGACTGATAGCATGACTAAAGGTTTGACGTTCAACAACAACGTTAAAGTAACATTGGATGGT


GAAGATTTTCCTGTTTTAAACTACAAACTCGTAACAGATGACCAAGGTTTCCGTCTTGCCTTGAATGCAACA


GGTCTTGCAGCAGTAGCAGCAGCTGCAAAAGACAAAGATGTTGAAATCAAGATCACTTACTCAGCTACGGTG


AACGGCTCCACTACTGTTGAAATTCCAGAAACCAATGATGTTAAATTGGACTATGGTAATAACCCAACGGAA


GAAAGTGAACCACAAGAAGGTACTCCAGCTAACCAAGAAATTAAAGTCATTAAAGACTGGGCAGTAGATGGT


ACAATTACTGATGCTAATGTTGCAGTTAAAGCTATCTTTACCTTGCAAGAAAAACAAACGGATGGTACATGG


GTGAACGTTGCTTCACACGAAGCAACAAAACCATCACGCTTTGAACATACTTTCACAGGTTTGGATAATGCT


AAAACTTACCGCGTTGTCGAACGTGTTAGCGGCTACACTCCAGAATACGTATCATTTAAAAATGGTGTTGTG


ACTATCAAGAACAACAAAAACTCAAATGATCCAACTCCAATCAACCCATCAGAACCAAAAGTGGTGACTTAT


GGACGTAAATTTGTGAAAACAAATCAAGCTAACACTGAACGCTTGGCAGGAGCTACCTTCCTCGTTAAGAAA


GAAGGCAAATACTTGGCACGTAAAGCAGGTGCAGCAACTGCTGAAGCAAAGGCAGCTGTAAAAACTGCTAAA


CTAGCATTGGATGAAGCTGTTAAAGCTTATAACGACTTGACTAAAGAAAAACAAGAAGGCCAAGAAGGTAAA


ACAGCATTGGCTACTGTTGATCAAAAACAAAAAGCTTACAATGACGCTTTTGTTAAAGCTAACTACTCATAT


GAATGGGTTGCAGATAAAAAGGCTGATAATGTTGTTAAATTGATCTCTAACGCCGGTGGTCAATTTGAAATT


ACTGGTTTGGATAAAGGCACTTATGGCTTGGAAGAAACTCAAGCACCAGCAGGTTATGCGACATTGTCAGGT


GATGTAAACTTTGAAGTAACTGCCACATCATATAGCAAAGGGGCTACAACTGACATCGCATATGATAAAGGC


TCTGTAAAAAAAGATGCCCAACAAGTTCAAAACAAAAAAGTAACCATCCCACAAACAGGTGGTATTGGTACA


ATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCAGTAGTTATCATGAAAAAACGTCAATCAGAG


GAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 114):


MKKINKCLTMFSTLLLILTSLFSVAPAFADDATTDTVTLHKIVMPQAAFDNFTEGTKGKNDSDYVGKQINDL


KSYFGSTDAKEIKGAFFVFKNETGTKFITENGKEVDTLEAKDAEGGAVLSGLTKDNGFVFNTAKLKGIYQIV


ELKEKSNYDNNGSILADSKAVPVKITLPLVNNQGVVKDAHIYPKNTETKPQVDKNFADKDLDYTDNRKDKGV


VSATVGDKKEYIVGTKILKGSDYKKLVWTDSMTKGLTFNNNVKVTLDGEDFPVLNYKLVTDDQGFRLALNAT


GLAAVAAAAKDKDVEIKITYSATVNGSTTVEIPETNDVKLDYGNNPTEESEPQEGTPANQEIKVIKDWAVDG


TITDANVAVKAIFTLQEKQTDGTWVNVASHEATKPSRFEHTFTGLDNAKTYRVVERVSGYTPEYVSFKNGVV


TIKNNKNSNDPTPINPSEPKVVTYGRKFVKTNQANTERLAGATFLVKKEGKYLARKAGAATAEAKAAVKTAK


LALDEAVKAYNDLTKEKQEGQEGKTALATVDQKQKAYNDAFVKANYSYEWVADKKADNVVKLISNAGGQFEI


TGLDKGTYGLEETQAPAGYATLSGDVNFEVTATSYSKGATTDIAYDKGSVKKDAQQVQNKKVTIPQTGGIGT


ILFTIIGLSIMLGAVVIMKKRQSEEA


 


Strain IC304


ORF DNA sequence (SEQ ID NO: 157):


ATGAAAAAAATCAACAAATGTCTTACAATGTTCTCGACACTGCTATTGATCTTAACGTCACTATTCTCAGTT


GCACCAGCGTTTGCGGACGACGCAACAACTGATACTGTGACCTTGCACAAGATTGTCATGCCACAAGCTGCA


TTTGATAACTTTACTGAAGGTACAAAAGGTAAGAATGATAGCGATTATGTTGGTAAACAAATTAATGACCTT


AAATCTTATTTTGGCTCAACCGATGCTAAAGAAATCAAGGGTGCTTTCTTTGTTTTCAAAAATGAAACTGGT


ACAAAATTCATTACTGAAAATGGTAAGGAAGTCGATACTTTGGAAGCTAAAGATGCTGAAGGTGGTGCTGTT


CTTTCAGGGTTAACAAAAGACAATGGTTTTGTTTTTAACACTGCTAAGTTAAAAGGAATTTACCAAATCGTT


GAATTGAAAGAAAAATCAAACTACGATAACAACGGTTCTATCTTGGCTGATTCAAAAGCAGTTCCAGTTAAA


ATCACTCTGCCATTGGTAAACAACCAAGGTGTTGTTAAAGATGCTCACATTTATCCAAAGAATACTGAAACA


AAACCACAAGTAGATAAGAACTTTGCAGATAAAGATCTTGATTATACTGACAACCGAAAAGACAAAGGTGTT


GTCTCAGCGACAGTTGGTGACAAAAAAGAATACATAGTTGGAACAAAAATTCTTAAAGGCTCAGACTATAAG


AAACTGGTTTGGACTGATAGCATGACTAAAGGTTTGACGTTCAACAACAACGTTAAAGTAACATTGGATGGT


GAAGATTTTCCTGTTTTAAACTACAAACTCGTAACAGATGACCAAGGTTTCCGTCTTGCCTTGAATGCAACA


GGTCTTGCAGCAGTAGCAGCAGCTGCAAAAGACAAAGATGTTGAAATCAAGATCACTTACTCAGCTACGGTG


AACGGCTCCACTACTGTTGAAATTCCAGAAACCAATGATGTTAAATTGGACTATGGTAATAACCCAACGGAA


GAAAGTGAACCACAAGAAGGTACTCCAGCTAACCAAGAAATTAAAGTCATTAAAGACTGGGCAGTAGATGGT


ACAATTACTGATGCTAATGTTGCAGTTAAAGCTATCTTTACCTTGCAAGAAAAACAAACGGATGGTACATGG


GTGAACGTTGCTTCACACGAAGCAACAAAACCATCACGCTTTGAACATACTTTCACAGGTTTGGATAATGCT


AAAACTTACCGCGTTGTCGAACGTGTTAGCGGCTACACTCCAGAATACGTATCATTTAAAAATGGTGTTGTG


ACTATCAAGAACAACAAAAACTCAAATGATCCAACTCCAATCAACCCATCAGAACCAAAAGTGGTGACTTAT


GGACGTAAATTTGTGAAAACAAATCAAGCTAACACTGAACGCTTGGCAGGAGCTACCTTCCTCGTTAAGAAA


GAAGGCAAATACTTGGCACGTAAAGCAGGTGCAGCAACTGCTGAAGCAAAGGCAGCTGTAAAAACTGCTAAA


CTAGCATTGGATGAAGCTGTTAAAGCTTATAACGACTTGACTAAAGAAAAACAAGAAGGCCAAGAAGGTAAA


ACAGCATTGGCTACTGTTGATCAAAAACAAAAAGCTTACAATGACGCTTTTGTTAAAGCTAACTACTCATAT


GAATGGGTTGCAGATAAAAAGGCTGATAATGTTGTTAAATTGATCTCTAACGCCGGTGGTCAATTTGAAATT


ACTGGTTTGGATAAAGGCACTTATGGCTTGGAAGAAACTCAAGCACCAGCAGGTTATGCGACATTGTCAGGT


GATGTAAACTTTGAAGTAACTGCCACATCATATAGCAAAGGGGCTACAACTGACATCGCATATGATAAAGGC


TCTGTAAAAAAAGATGCCCAACAAGTTCAAAACAAAAAAGTAACCATCCCACAAACAGGTGGTATTGGTACA


ATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCAGTAGTTATCATGAAAAAACGTCAATCAGAG


GAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 115):


MKKINKCLTMFSTLLLILTSLFSVAPAFADDATTDTVTLHKIVMPQAAFDNFTEGTKGKNDSDYVGKQINDL


KSYFGSTDAKEIKGAFFVFKNETGTKFITENGKEVDTLEAKDAEGGAVLSGLTKDNGFVFNTAKLKGIYQIV


ELKEKSNYDNNGSILADSKAVPVKITLPLVNNQGVVKDAHIYPKNTETKPQVDKNFADKDLDYTDNRKDKGV


VSATVGDKKEYIVGTKILKGSDYKKLVWTDSMTKGLTFNNNVKVTLDGEDFPVLNYKLVTDDQGFRLALNAT


GLAAVAAAAKDKDVEIKITYSATVNGSTTVEIPETNDVKLDYGNNPTEESEPQEGTPANQEIKVIKDWAVDG


TITDANVAVKAIFTLQEKQTDGTWVNVASHEATKPSRFEHTFTGLDNAKTYRVVERVSGYTPEYVSFKNGVV


TIKNNKNSNDPTPINPSEPKVVTYGRKFVKTNQANTERLAGATFLVKKEGKYLARKAGAATAEAKAAVKTAK


LALDEAVKAYNDLTKEKQEGQEGKTALATVDQKQKAYNDAFVKANYSYEWVADKKADNVVKLISNAGGQFEI


TGLDKGTYGLEETQAPAGYATLSGDVNFEVTATSYSKGATTDIAYDKGSVKKDAQQVQNKKVTIPQTGGIGT


ILFTIIGLSIMLGAVVIMKKRQSEEA


 


Strain IC305


ORF DNA sequence (SEQ ID NO: 158):


ATGAAAAAAATCAACAAATGTCTTACAATGTTCTCGACACTGCTATTGATCTTAACGTCACTATTCTCAGTT


GCACCAGCGTTTGCGGACGACGCAACAACTGATACTGTGACCTTGCACAAGATTGTCATGCCACAAGCTGCA


TTTGATAACTTTACTGAAGGTACAAAAGGTAAGAATGATAGCGATTATGTTGGTAAACAAATTAATGACCTT


AAATCTTATTTTGGCTCAACCGATGCTAAAGAAATCAAGGGTGCTTTCTTTGTTTTCAAAAATGAAACTGGT


ACAAAATTCATTACTGAAAATGGTAAGGAAGTCGATACTTTGGAAGCTAAAGATGCTGAAGGTGGTGCTGTT


CTTTCAGGGTTAACAAAAGACAATGGTTTTGTTTTTAACACTGCTAAGTTAAAAGGAATTTACCAAATCGTT


GAATTGAAAGAAAAATCAAACTACGATAACAACGGTTCTATCTTGGCTGATTCAAAAGCAGTTCCAGTTAAA


ATCACTCTGCCATTGGTAAACAACCAAGGTGTTGTTAAAGATGCTCACATTTATCCAAAGAATACTGAAACA


AAACCACAAGTAGATAAGAACTTTGCAGATAAAGATCTTGATTATACTGACAACCGAAAAGACAAAGGTGTT


GTCTCAGCGACAGTTGGTGACAAAAAAGAATACATAGTTGGAACAAAAATTCTTAAAGGCTCAGACTATAAG


AAACTGGTTTGGACTGATAGCATGACTAAAGGTTTGACGTTCAACAACAACGTTAAAGTAACATTGGATGGT


GAAGATTTTCCTGTTTTAAACTACAAACTCGTAACAGATGACCAAGGTTTCCGTCTTGCCTTGAATGCAACA


GGTCTTGCAGCAGTAGCAGCAGCTGCAAAAGACAAAGATGTTGAAATCAAGATCACTTACTCAGCTACGGTG


AACGGCTCCACTACTGTTGAAATTCCAGAAACCAATGATGTTAAATTGGACTATGGTAATAACCCAACGGAA


GAAAGTGAACCACAAGAAGGTACTCCAGCTAACCAAGAAATTAAAGTCATTAAAGACTGGGCAGTAGATGGT


ACAATTACTGATGCTAATGTTGCAGTTAAAGCTATCTTTACCTTGCAAGAAAAACAAACGGATGGTACATGG


GTGAACGTTGCTTCACACGAAGCAACAAAACCATCACGCTTTGAACATACTTTCACAGGTTTGGATAATGCT


AAAACTTACCGCGTTGTCGAACGTGTTAGCGGCTACACTCCAGAATACGTATCATTTAAAAATGGTGTTGTG


ACTATCAAGAACAACAAAAACTCAAATGATCCAACTCCAATCAACCCATCAGAACCAAAAGTGGTGACTTAT


GGACGTAAATTTGTGAAAACAAATCAAGCTAACACTGAACGCTTGGCAGGAGCTACCTTCCTCGTTAAGAAA


GAAGGCAAATACTTGGCACGTAAAGCAGGTGCAGCAACTGCTGAAGCAAAGGCAGCTGTAAAAACTGCTAAA


CTAGCATTGGATGAAGCTGTTAAAGCTTATAACGACTTGACTAAAGAAAAACAAGAAGGCCAAGAAGGTAAA


ACAGCATTGGCTACTGTTGATCAAAAACAAAAAGCTTACAATGACGCTTTTGTTAAAGCTAACTACTCATAT


GAATGGGTTGCAGATAAAAAGGCTGATAATGTTGTTAAATTGATCTCTAACGCCGGTGGTCAATTTGAAATT


ACTGGTTTGGATAAAGGCACTTATGGCTTGGAAGAAACTCAAGCACCAGCAGGTTATGCGACATTGTCAGGT


GATGTAAACTTTGAAGTAACTGCCACATCATATAGCAAAGGGGCTACAACTGACATCGCATATGATAAAGGC


TCTGTAAAAAAAGATGCCCAACAAGTTCAAAACAAAAAAGTAACCATCCCACAAACAGGTGGTATTGGTACA


ATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCAGTAGTTATCATGAAAAAACGTCAATCAGAG


GAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 116):


MKKINKCLTMFSTLLLILTSLFSVAPAFADDATTDTVTLHKIVMPQAAFDNFTEGTKGKNDSDYVGKQINDL


KSYFGSTDAKEIKGAFFVFKNETGTKFITENGKEVDTLEAKDAEGGAVLSGLTKDNGFVFNTAKLKGIYQIV


ELKEKSNYDNNGSILADSKAVPVKITLPLVNNQGVVKDAHIYPKNTETKPQVDKNFADKDLDYTDNRKDKGV


VSATVGDKKEYIVGTKILKGSDYKKLVWTDSMTKGLTFNNNVKVTLDGEDFPVLNYKLVTDDQGFRLALNAT


GLAAVAAAAKDKDVEIKITYSATVNGSTTVEIPETNDVKLDYGNNPTEESEPQEGTPANQEIKVIKDWAVDG


TITDANVAVKAIFTLQEKQTDGTWVNVASHEATKPSRFEHTFTGLDNAKTYRVVERVSGYTPEYVSFKNGVV


TIKNNKNSNDPTPINPSEPKVVTYGRKFVKTNQANTERLAGATFLVKKEGKYLARKAGAATAEAKAAVKTAK


LALDEAVKAYNDLTKEKQEGQEGKTALATVDQKQKAYNDAFVKANYSYEWVADKKADNVVKLISNAGGQFEI


TGLDKGTYGLEETQAPAGYATLSGDVNFEVTATSYSKGATTDIAYDKGSVKKDAQQVQNKKVTIPQTGGIGT


ILFTIIGLSIMLGAVVIMKKRQSEEA


 


Strain IC306


ORF DNA sequence (SEQ ID NO: 159):


ATGAAAAAAATCAACAAATGTCTTACAATGTTCTCGACACTGCTATTGATCTTAACGTCACTATTCTCAGTT


GCACCAGCGTTTGCGGACGACGCAACAACTGATACTGTGACCTTGCACAAGATTGTCATGCCACAAGCTGCA


TTTGATAACTTTACTGAAGGTACAAAAGGTAAGAATGATAGCGATTATGTTGGTAAACAAATTAATGACCTT


AAATCTTATTTTGGCTCAACCGATGCTAAAGAAATCAAGGGTGCTTTCTTTGTTTTCAAAAATGAAACTGGT


ACAAAATTCATTACTGAAAATGGTAAGGAAGTCGATACTTTGGAAGCTAAAGATGCTGAAGGTGGTGCTGTT


CTTTCAGGGTTAACAAAAGACAATGGTTTTGTTTTTAACACTGCTAAGTTAAAAGGAATTTACCAAATCGTT


GAATTGAAAGAAAAATCAAACTACGATAACAACGGTTCTATCTTGGCTGATTCAAAAGCAGTTCCAGTTAAA


ATCACTCTGCCATTGGTAAACAACCAAGGTGTTGTTAAAGATGCTCACATTTATCCAAAGAATACTGAAACA


AAACCACAAGTAGATAAGAACTTTGCAGATAAAGATCTTGATTATACTGACAACCGAAAAGACAAAGGTGTT


GTCTCAGCGACAGTTGGTGACAAAAAAGAATACATAGTTGGAACAAAAATTCTTAAAGGCTCAGACTATAAG


AAACTGGTTTGGACTGATAGCATGACTAAAGGTTTGACGTTCAACAACAACGTTAAAGTAACATTGGATGGT


GAAGATTTTCCTGTTTTAAACTACAAACTCGTAACAGATGACCAAGGTTTCCGTCTTGCCTTGAATGCAACA


GGTCTTGCAGCAGTAGCAGCAGCTGCAAAAGACAAAGATGTTGAAATCAAGATCACTTACTCAGCTACGGTG


AACGGCTCCACTACTGTTGAAATTCCAGAAACCAATGATGTTAAATTGGACTATGGTAATAACCCAACGGAA


GAAAGTGAACCACAAGAAGGTACTCCAGCTAACCAAGAAATTAAAGTCATTAAAGACTGGGCAGTAGATGGT


ACAATTACTGATGCTAATGTTGCAGTTAAAGCTATCTTTACCTTGCAAGAAAAACAAACGGATGGTACATGG


GTGAACGTTGCTTCACACGAAGCAACAAAACCATCACGCTTTGAACATACTTTCACAGGTTTGGATAATGCT


AAAACTTACCGCGTTGTCGAACGTGTTAGCGGCTACACTCCAGAATACGTATCATTTAAAAATGGTGTTGTG


ACTATCAAGAACAACAAAAACTCAAATGATCCAACTCCAATCAACCCATCAGAACCAAAAGTGGTGACTTAT


GGACGTAAATTTGTGAAAACAAATCAAGCTAACACTGAACGCTTGGCAGGAGCTACCTTCCTCGTTAAGAAA


GAAGGCAAATACTTGGCACGTAAAGCAGGTGCAGCAACTGCTGAAGCAAAGGCAGCTGTAAAAACTGCTAAA


CTAGCATTGGATGAAGCTGTTAAAGCTTATAACGACTTGACTAAAGAAAAACAAGAAGGCCAAGAAGGTAAA


ACAGCATTGGCTACTGTTGATCAAAAACAAAAAGCTTACAATGACGCTTTTGTTAAAGCTAACTACTCATAT


GAATGGGTTGCAGATAAAAAGGCTGATAATGTTGTTAAATTGATCTCTAACGCCGGTGGTCAATTTGAAATT


ACTGGTTTGGATAAAGGCACTTATGGCTTGGAAGAAACTCAAGCACCAGCAGGTTATGCGACATTGTCAGGT


GATGTAAACTTTGAAGTAACTGCCACATCATATAGCAAAGGGGCTACAACTGACATCGCATATGATAAAGGC


TCTGTAAAAAAAGATGCCCAACAAGTTCAAAACAAAAAAGTAACCATCCCACAAACAGGTGGTATTGGTACA


ATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCAGTAGTTATCATGAAAAAACGTCAATCAGAG


GAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 117):


MKKINKCLTMFSTLLLILTSLFSVAPAFADDATTDTVTLHKIVMPQAAFDNFTEGTKGKNDSDYVGKQINDL


KSYFGSTDAKEIKGAFFVFKNETGTKFITENGKEVDTLEAKDAEGGAVLSGLTKDNGFVFNTAKLKGIYQIV


ELKEKSNYDNNGSILADSKAVPVKITLPLVNNQGVVKDAHIYPKNTETKPQVDKNFADKDLDYTDNRKDKGV


VSATVGDKKEYIVGTKILKGSDYKKLVWTDSMTKGLTFNNNVKVTLDGEDFPVLNYKLVTDDQGFRLALNAT


GLAAVAAAAKDKDVEIKITYSATVNGSTTVEIPETNDVKLDYGNNPTEESEPQEGTPANQEIKVIKDWAVDG


TITDANVAVKAIFTLQEKQTDGTWVNVASHEATKPSRFEHTFTGLDNAKTYRVVERVSGYTPEYVSFKNGVV


TIKNNKNSNDPTPINPSEPKVVTYGRKFVKTNQANTERLAGATFLVKKEGKYLARKAGAATAEAKAAVKTAK


LALDEAVKAYNDLTKEKQEGQEGKTALATVDQKQKAYNDAFVKANYSYEWVADKKADNVVKLISNAGGQFEI


TGLDKGTYGLEETQAPAGYATLSGDVNFEVTATSYSKGATTDIAYDKGSVKKDAQQVQNKKVTIPQTGGIGT


ILFTIIGLSIMLGAVVIMKKRQSEEA


 


Strain IC361


ORF DNA sequence (SEQ ID NO: 160):


ATGAAAAAAATCAACAAATGTCTTACAATGTTCTCGACACTGCTATTGATCTTAACGTCACTATTCTCAGTT


GCACCAGCGTTTGCGGACGACGCAACAACTGATACTGTGACCTTGCACAAGATTGTCATGCCACAAGCTGCA


TTTGATAACTTTACTGAAGGTACAAAAGGTAAGAATGATAGCGATTATGTTGGTAAACAAATTAATGACCTT


AAATCTTATTTTGGCTCAACCGATGCTAAAGAAATCAAGGGTGCTTTCTTTGTTTTCAAAAATGAAACTGGT


ACAAAATTCATTACTGAAAATGGTAAGGAAGTCGATACTTTGGAAGCTAAAGATGCTGAAGGTGGTGCTGTT


CTTTCAGGGTTAACAAAAGACAATGGTTTTGTTTTTAACACTGCTAAGTTAAAAGGAATTTACCAAATCGTT


GAATTGAAAGAAAAATCAAACTACGATAACAACGGTTCTATCTTGGCTGATTCAAAAGCAGTTCCAGTTAAA


ATCACTCTGCCATTGGTAAACAACCAAGGTGTTGTTAAAGATGCTCACATTTATCCAAAGAATACTGAAACA


AAACCACAAGTAGATAAGAACTTTGCAGATAAAGATCTTGATTATACTGACAACCGAAAAGACAAAGGTGTT


GTCTCAGCGACAGTTGGTGACAAAAAAGAATACATAGTTGGAACAAAAATTCTTAAAGGCTCAGACTATAAG


AAACTGGTTTGGACTGATAGCATGACTAAAGGTTTGACGTTCAACAACAACGTTAAAGTAACATTGGATGGT


GAAGATTTTCCTGTTTTAAACTACAAACTCGTAACAGATGACCAAGGTTTCCGTCTTGCCTTGAATGCAACA


GGTCTTGCAGCAGTAGCAGCAGCTGCAAAAGACAAAGATGTTGAAATCAAGATCACTTACTCAGCTACGGTG


AACGGCTCCACTACTGTTGAAATTCCAGAAACCAATGATGTTAAATTGGACTATGGTAATAACCCAACGGAA


GAAAGTGAACCACAAGAAGGTACTCCAGCTAACCAAGAAATTAAAGTCATTAAAGACTGGGCAGTAGATGGT


ACAATTACTGATGCTAATGTTGCAGTTAAAGCTATCTTTACCTTGCAAGAAAAACAAACGGATGGTACATGG


GTGAACGTTGCTTCACACGAAGCAACAAAACCATCACGCTTTGAACATACTTTCACAGGTTTGGATAATGCT


AAAACTTACCGCGTTGTCGAACGTGTTAGCGGCTACACTCCAGAATACGTATCATTTAAAAATGGTGTTGTG


ACTATCAAGAACAACAAAAACTCAAATGATCCAACTCCAATCAACCCATCAGAACCAAAAGTGGTGACTTAT


GGACGTAAATTTGTGAAAACAAATCAAGCTAACACTGAACGCTTGGCAGGAGCTACCTTCCTCGTTAAGAAA


GAAGGCAAATACTTGGCACGTAAAGCAGGTGCAGCAACTGCTGAAGCAAAGGCAGCTGTAAAAACTGCTAAA


CTAGCATTGGATGAAGCTGTTAAAGCTTATAACGACTTGACTAAAGAAAAACAAGAAGGCCAAGAAGGTAAA


ACAGCATTGGCTACTGTTGATCAAAAACAAAAAGCTTACAATGACGCTTTTGTTAAAGCTAACTACTCATAT


GAATGGGTTGCAGATAAAAAGGCTGATAATGTTGTTAAATTGATCTCTAACGCCGGTGGTCAATTTGAAATT


ACTGGTTTGGATAAAGGCACTTATGGCTTGGAAGAAACTCAAGCACCAGCAGGTTATGCGACATTGTCAGGT


GATGTAAACTTTGAAGTAACTGCCACATCATATAGCAAAGGGGCTACAACTGACATCGCATATGATAAAGGC


TCTGTAAAAAAAGATGCCCAACAAGTTCAAAACAAAAAAGTAACCATCCCACAAACAGGTGGTATTGGTACA


ATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCAGTAGTTATCATGAAAAAACGTCAATCAGAG


GAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 118):


MKKINKCLTMFSTLLLILTSLFSVAPAFADDATTDTVTLHKIVMPQAAFDNFTEGTKGKNDSDYVGKQINDL


KSYFGSTDAKEIKGAFFVFKNETGTKFITENGKEVDTLEAKDAEGGAVLSGLTKDNGFVFNTAKLKGIYQIV


ELKEKSNYDNNGSILADSKAVPVKITLPLVNNQGVVKDAHIYPKNTETKPQVDKNFADKDLDYTDNRKDKGV


VSATVGDKKEYIVGTKILKGSDYKKLVWTDSMTKGLTFNNNVKVTLDGEDFPVLNYKLVTDDQGFRLALNAT


GLAAVAAAAKDKDVEIKITYSATVNGSTTVEIPETNDVKLDYGNNPTEESEPQEGTPANQEIKVIKDWAVDG


TITDANVAVKAIFTLQEKQTDGTWVNVASHEATKPSRFEHTFTGLDNAKTYRVVERVSGYTPEYVSFKNGVV


TIKNNKNSNDPTPINPSEPKVVTYGRKFVKTNQANTERLAGATFLVKKEGKYLARKAGAATAEAKAAVKTAK


LALDEAVKAYNDLTKEKQEGQEGKTALATVDQKQKAYNDAFVKANYSYEWVADKKADNVVKLISNAGGQFEI


TGLDKGTYGLEETQAPAGYATLSGDVNFEVTATSYSKGATTDIAYDKGSVKKDAQQVQNKKVTIPQTGGIGT


ILFTIIGLSIMLGAVVIMKKRQSEEA


 


Strain IC363


ORF DNA sequence (SEQ ID NO: 161):


ATGAAAAGAATCAACAAATATTTTGCAATGTTCTCGGCATTGTTACTGACTTTAACGTCATTGCTCTCAGTT


GCACCAGCGTTTGCGGATGAAGCAACAACTAATACAGTGACTTTGCACAAGATTTTGCAAACCGAATCAAAT


CTTAACAAAAGTAACTTCCCAGGAACTACAGGTCTTAACGGAAAAGACTACAAAGGTGGAGCTATTTCTGAC


CTTGCTGGTTACTTTGGCGAGGGATCTAAAGAAATCGAAGGTGCGTTCTTTGCTTTAGCTTTGAAAGAAGAT


AAAAGTGGTAAAGTGCAATATGTTAAGGCAAAAGAAGGTAACAAATTAACACCAGCCTTAATTAATAAAGAT


GGTACTCCTGAAATAACAGTAAATATTGATGAGGCCGTGTCTGGATTGACACCAGAGGGAGATACTGGACTT


GTTTTCAACACCAAAGGATTGAAAGGCGAGTTTAAAATTGTTGAAGTTAAATCAAAATCTACTTACAACAAT


AATGGTTCCCTCCTGGCTGCTTCAAAAGCGGTTCCAGTTAACATCACTCTTCCATTGGTAAATGAAGATGGT


GTTGTTGCTGATGCCCATGTTTATCCAAAGAACACTGAAGAAAAACCAGAAATTGATAAAAACTTTGCTAAA


ACAAACGATTTGACAGCATTGACAGATGTTAATAGACTTTTGACAGCTGGCGCAAATTATGGTAATTATGCA


CGTGACAAAGCAACTGCTACTGCTGAAATCGGTAAAGTTGTTCCTTATGAAGTTAAAACAAAAATTCACAAA


GGTTCTAAATACGAAAACTTGGTTTGGACAGATATAATGTCAAATGGTTTGACAATGGGTTCAACTGTTAGC


CTTAAAGCTTCAGGAACTACAGAAACTTTTGCTAAGGATACAGACTATGAACTTAGCATTGATGCCCGTGGT


TTCACATTAAAATTCACAGCTGATGGATTGGGCAAATTGGAAAAAGCAGCTAAAACAGCTGATATTGAATTT


ACATTGACTTATAGTGCTACTGTTAATGGTCAAGCAATTATTGATAATCCAGAATCCAATGATATCAAATTG


TCGTATGGTAACAAACCAGGTAAAGACTTGACTGAACTTCCTGTTACACCTTCAAAGGGTGAAGTAACAGTT


GCTAAAACTTGGTCTGACGGAATTGCACCTGATGGTGTAAACGTTGTTTACACATTGAAAGATAAAGATAAA


ACTGTTGCTTCAGTATCATTGACAAAAACATCTAAAGGTACAATCGACCTTGGAAATGGTATCAAATTTGAA


GTATCTGGTAACTTCTCGGGTAAATTCACTGGTCTAGAAAACAAATCATACATGATCTCAGAACGTGTTTCT


GGTTACGGAAGTGCAATAAATCTAGAAAATGGTAAAGTAACCATTACCAATACCAAAGATTCTGATAACCCA


ACACCATTGAACCCAACTGAACCAAAAGTTGAAACTCATGGTAAGAAATTTGTCAAAACTAATGAACAAGGT


GACCGTTTGGCTGGTGCACAATTCGTTGTGAAAAACTCAGCAGGTAAATACCTTGCTCTTAAAGCAGATCAA


TCAGAAGGTCAAAAAACTTTAGCTGCTAAGAAAATAGCTTTAGATGAAGCTATCGCTGCTTATAACAAGTTG


TCTGCAACAGACCAAAAAGGTGAAAAAGGAATTACTGCAAAAGAACTTATCAAAACTAAACAAGCAGATTAC


GATGCAGCCTTCATTGAGGCTCGTACAGCTTATGAGTGGATAACAGATAAGGCTAGAGCCATTACCTACACT


TCAAACGATCAAGGTCAATTTGAAGTTACAGGTCTTGCAGACGGTACTTACAACCTTGAAGAAACACTTGCT


CCAGCAGGATTTGCTAAGTTGGCAGGTAATATTAAGTTTGTAGTTAATCAAGGGTCATACATAACAGGTGGT


AACATTGACTACGTTGCTAACAGCAACCAAAAAGATGCGACACGTGTAGAAAATAAAAAGGTAACAATCCCA


CAAACAGGTGGTATTGGTACAATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCAGTAGTTATC


ATGAAAAGACGCCAATCAAAGGAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 119):


MKRINKYFAMFSALLLTLTSLLSVAPAFADEATTNTVTLHKILQTESNLNKSNFPGTTGLNGKDYKGGAISD


LAGYFGEGSKEIEGAFFALALKEDKSGKVQYVKAKEGNKLTPALINKDGTPEITVNIDEAVSGLTPEGDTGL


VFNTKGLKGEFKIVEVKSKSTYNNNGSLLAASKAVPVNITLPLVNEDGVVADAHVYPKNTEEKPEIDKNFAK


TNDLTALTDVNRLLTAGANYGNYARDKATATAEIGKVVPYEVKTKIHKGSKYENLVWTDIMSNGLTMGSTVS


LKASGTTETFAKDTDYELSIDARGFTLKFTADGLGKLEKAAKTADIEFTLTYSATVNGQAIIDNPESNDIKL


SYGNKPGKDLTELPVTPSKGEVTVAKTWSDGIAPDGVNVVYTLKDKDKTVASVSLTKTSKGTIDLGNGIKFE


VSGNFSGKFTGLENKSYMISERVSGYGSAINLENGKVTITNTKDSDNPTPLNPTEPKVETHGKKFVKTNEQG


DRLAGAQFVVKNSAGKYLALKADQSEGQKTLAAKKIALDEATAAYNKLSATDQKGEKGITAKELIKTKQADY


DAAFIEARTAYEWITDKARAITYTSNDQGQFEVTGLADGTYNLEETLAPAGFAKLAGNIKFVVNQGSYITGG


NIDYVANSNQKDATRVENKKVTIPQTGGIGTILFTIIGLSIMLGAVVIMKRRQSKEA


 


Strain IC365


ORF DNA sequence (SEQ ID NO: 162):


ATGAAAAAAATCAACAAATATTTTGCAGTCTTCTCGGCATTGCTACTGACCGTAACATCATTGTTCTCAGTT


GCACCAGTGTTTGCGGAAGAAGCAAAAACTACTGACACAGTGACCTTGCACAAGATTGTCATGCCTCGAACT


GCATTTGACGGTTTTACTGCTGGTACAAAGGGTAAGGATAATACTGACTACGTTGGTAAACAAATCGAAGAC


CTTAAAACTTACTTTGGCTCAGGCGAAGCGAAAGAAATCGCAGGTGCTTACTTTGCTTTCAAAAATGAAGCT


GGTACTAAATACATCACTGAAAATGGTGAAGAAGTTGATACTTTGGATACAACAGATGCCAAAGGTTGTGCT


GTTCTTAAAGGTTTAACAACAGACAATGGTTTCAAATTTAACACTTCTAAATTAACAGGAACTTACCAAATC


GTTGAATTGAAAGAAAAATCTACATACAACAACGATGGTTCTATCTTGGCTGATTCAAAAGCAGTTCCAGTT


AAAATCACTCTTCCATTGGTAAACGACAATGGTGTTGTTAAAGACGCTCACGTTTATCCAAAGAACACTGAA


ACAAAACCACAAGTAGATAAGAACTTCGCAGATAAAGAACTTGATTATGCGAACAACAAAAAAGACAAAGGG


ACTGTCTCAGCATCTGTTGGTGATGTTAAAAAATACCATGTTGGAACAAAAATCCTTAAAGGTTCAGACTAT


AAGAAATTAATCTGGACCGATAGCATGACCAAAGGTTTGACTTTCAACAACGATATTGCTGTAACATTGGAT


GGTGCAACTCTTGATGCTACAAATTACAAACTTGTAGCAGATGACCAAGGTTTCCGCCTTGTCTTGACTGAC


AAAGGTCTTGAAGCAGTGGCAAAAGCCGCAAAAACAAAAGATGTTGAAATCAAGATCACTTACTCAGCTACT


TTGAACGGTTCTGCTGTCGTTGAAGTTCTAGAAACCAATGATGTTAAATTGGACTACGGCAACAACCCAACA


ATTGAAAATGAACCAAAAGAAGGTATTCCAGTTGATAAGAAAATCACTGTTAACAAAACATGGGCAGTAGAT


GGCAATGAAGTGAATAAAGCAGATGAAACAGTTGATGCTGTCTTCACCTTGCAAGTTAAAGATGGTGACAAA


TGGGTGAATGTTGATTCAGCTAAAGCAACAGCTGCAACTAGCTTCAAACACACTTTTGAAAACTTGGATAAT


GCTAAAACTTACCGCGTTATCGAACGTGTTAGCGGCTACGCTCCAGAATACGTCTCATTTGTAAATGGCGTT


GTAACCATCAAGAACAACAAAGACTCAAATGAGCCAACTCCAATCAACCCATCAGAACCAAAAGTGGTGACT


TATGGACGTAAATTTGTGAAAACAAATAAAGATGGAAAAGAACGCTTGGCAGGAGCTACCTTCCTTGTTAAG


AAAGATGGCAAGTACTTGGCACGTAAATCAGGTGTTGCAACAGATGCAGAAAAAGCTGCTGTAGATTCAACT


AAATCAGCATTGGATGCTGCTGTTAAAGCTTACAATGATTTGACTAAAGAAAAACAAGAAGGTCAAGATGGT


AAATCAGCATTGGCTACCGTTAGTGAAAAACAAAAAGCTTACAATGATGCCTTTGTTAAAGCTAACTACTCA


TACGAATGGGTTGAAGATAAAAATGCTAAGAATGTTGTTAAATTGATTTCTAACGATAAAGGTCAATTTGAA


ATTACTGGCTTGACTGAAGGTCAATACTCATTGGAAGAAACACAAGCACCAACTGGTTATGCTAAATTATCA


GGTGATGTTTCGTTTAATGTTAATGCTACTTCATACAGTAAAGGTTCTGCTCAAGATATTGAGTATACCCAA


GGTTCTAAAACTAAAGATGCACAACAAGTTATCAATAAGAAGGTTACTATTCCACAAACAGGTGGTATTGGT


ACAATTTTTTTCACAATTATTGGATTAAGTATTATGCTTGGAGCGGTAGTTATCATGAAAAGACGTCAATCA


GAGGAAGTTTAA


 


ORF amino acid sequence (SEQ ID NO: 120):


MKKINKYFAVFSALLLTVTSLFSVAPVFAEEAKTTDTVTLHKIVMPRTAFDGFTAGTKGKDNTDYVGKQIED


LKTYFGSGEAKEIAGAYFAFKNEAGTKYITENGEEVDTLDTTDAKGCAVLKGLTTDNGFKFNTSKLTGTYQI


VELKEKSTYNNDGSILADSKAVPVKITLPLVNDNGVVKDAHVYPKNTETKPQVDKNFADKELDYANNKKDKG


TVSASVGDVKKYHVGTKILKGSDYKKLIWTDSMTKGLTFNNDIAVTLDGATLDATNYKLVADDQGFRLVLTD


KGLEAVAKAAKTKDVEIKITYSATLNGSAVVEVLETNDVKLDYGNNPTIENEPKEGIPVDKKITVNKTWAVD


GNEVNKADETVDAVFTLQVKDGDKWVNVDSAKATAATSFKHTFENLDNAKTYRVIERVSGYAPEYVSFVNGV


VTIKNNKDSNEPTPINPSEPKVVTYGRKFVKTNKDGKERLAGATFLVKKDGKYLARKSGVATDAEKAAVDST


KSALDAAVKAYNDLTKEKQEGQDGKSALATVSEKQKAYNDAFVKANYSYEWVEDKNAKNVVKLISNDKGQFE


ITGLTEGQYSLEETQAPTGYAKLSGDVSFNVNATSYSKGSAQDIEYTQGSKTKDAQQVINKKVTIPQTGGIG


TIFFTIIGLSIMLGAVVIMKRRQSEEV


 


Strain IC367


ORF DNA sequence (SEQ ID NO: 163):


ATGAAAAAAATCAACAAATGTCTTACAATGTTCTCGACACTGCTATTGATCTTAACGTCACTATTCTCAGTT


GCACCAGCGTTTGCGGACGACGCAACAACTGATACTGTGACCTTGCACAAGATTGTCATGCCACAAGCTGCA


TTTGATAACTTTACTGAAGGTACAAAAGGTAAGAATGATAGCGATTATGTTGGTAAACAAATTAATGACCTT


AAATCTTATTTTGGCTCAACCGATGCTAAAGAAATCAAGGGTGCTTTCTTTGTTTTCAAAAATGAAACTGGT


ACAAAATTCATTACTGAAAATGGTAAGGAAGTCGATACTTTGGAAGCTAAAGATGCTGAAGGTGGTGCTGTT


CTTTCAGGGTTAACAAAAGACAATGGTTTTGTTTTTAACACTGCTAAGTTAAAAGGAATTTACCAAATCGTT


GAATTGAAAGAAAAATCAAACTACGATAACAACGGTTCTATCTTGGCTGATTCAAAAGCAGTTCCAGTTAAA


ATCACTCTGCCATTGGTAAACAACCAAGGTGTTGTTAAAGATGCTCACATTTATCCAAAGAATACTGAAACA


AAACCACAAGTAGATAAGAACTTTGCAGATAAAGATCTTGATTATACTGACAACCGAAAAGACAAAGGTGTT


GTCTCAGCGACAGTTGGTGACAAAAAAGAATACATAGTTGGAACAAAAATTCTTAAAGGCTCAGACTATAAG


AAACTGGTTTGGACTGATAGCATGACTAAAGGTTTGACGTTCAACAACAACGTTAAAGTAACATTGGATGGT


GAAGATTTTCCTGTTTTAAACTACAAACTCGTAACAGATGACCAAGGTTTCCGTCTTGCCTTGAATGCAACA


GGTCTTGCAGCAGTAGCAGCAGCTGCAAAAGACAAAGATGTTGAAATCAAGATCACTTACTCAGCTACGGTG


AACGGCTCCACTACTGTTGAAATTCCAGAAACCAATGATGTTAAATTGGACTATGGTAATAACCCAACGGAA


GAAAGTGAACCACAAGAAGGTACTCCAGCTAACCAAGAAATTAAAGTCATTAAAGACTGGGCAGTAGATGGT


ACAATTACTGATGCTAATGTTGCAGTTAAAGCTATCTTTACCTTGCAAGAAAAACAAACGGATGGTACATGG


GTGAACGTTGCTTCACACGAAGCAACAAAACCATCACGCTTTGAACATACTTTCACAGGTTTGGATAATGCT


AAAACTTACCGCGTTGTCGAACGTGTTAGCGGCTACACTCCAGAATACGTATCATTTAAAAATGGTGTTGTG


ACTATCAAGAACAACAAAAACTCAAATGATCCAACTCCAATCAACCCATCAGAACCAAAAGTGGTGACTTAT


GGACGTAAATTTGTGAAAACAAATCAAGCTAACACTGAACGCTTGGCAGGAGCTACCTTCCTCGTTAAGAAA


GAAGGCAAATACTTGGCACGTAAAGCAGGTGCAGCAACTGCTGAAGCAAAGGCAGCTGTAAAAACTGCTAAA


CTAGCATTGGATGAAGCTGTTAAAGCTTATAACGACTTGACTAAAGAAAAACAAGAAGGCCAAGAAGGTAAA


ACAGCATTGGCTACTGTTGATCAAAAACAAAAAGCTTACAATGACGCTTTTGTTAAAGCTAACTACTCATAT


GAATGGGTTGCAGATAAAAAGGCTGATAATGTTGTTAAATTGATCTCTAACGCCGGTGGTCAATTTGAAATT


ACTGGTTTGGATAAAGGCACTTATGGCTTGGAAGAAACTCAAGCACCAGCAGGTTATGCGACATTGTCAGGT


GATGTAAACTTTGAAGTAACTGCCACATCATATAGCAAAGGGGCTACAACTGACATCGCATATGATAAAGGC


TCTGTAAAAAAAGATGCCCAACAAGTTCAAAACAAAAAAGTAACCATCCCACAAACAGGTGGTATTGGTACA


ATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCAGTAGTTATCATGAAAAAACGTCAATCAGAG


GAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 121):


MKKINKCLTMFSTLLLILTSLFSVAPAFADDATTDTVTLHKIVMPQAAFDNFTEGTKGKNDSDYVGKQINDL


KSYFGSTDAKEIKGAFFVFKNETGTKFITENGKEVDTLEAKDAEGGAVLSGLTKDNGFVFNTAKLKGIYQIV


ELKEKSNYDNNGSILADSKAVPVKITLPLVNNQGVVKDAHIYPKNTETKPQVDKNFADKDLDYTDNRKDKGV


VSATVGDKKEYIVGTKILKGSDYKKLVWTDSMTKGLTFNNNVKVTLDGEDFPVLNYKLVTDDQGFRLALNAT


GLAAVAAAAKDKDVEIKITYSATVNGSTTVEIPETNDVKLDYGNNPTEESEPQEGTPANQEIKVIKDWAVDG


TITDANVAVKAIFTLQEKQTDGTWVNVASHEATKPSRFEHTFTGLDNAKTYRVVERVSGYTPEYVSFKNGVV


TIKNNKNSNDPTPINPSEPKVVTYGRKFVKTNQANTERLAGATFLVKKEGKYLARKAGAATAEAKAAVKTAK


LALDEAVKAYNDLTKEKQEGQEGKTALATVDQKQKAYNDAFVKANYSYEWVADKKADNVVKLISNAGGQFEI


TGLDKGTYGLEETQAPAGYATLSGDVNFEVTATSYSKGATTDIAYDKGSVKKDAQQVQNKKVTIPQTGGIGT


ILFTIIGLSIMLGAVVIMKKRQSEEA


 


Strain IC377


ORF DNA sequence (SEQ ID NO: 164):


ATGAAAAAAATCAACAAATGTCTTACAATGTTCTCGACACTGCTATTGATCTTAACGTCACTATTCTCAGTT


GCACCAGCGTTTGCGGACGACGCAACAACTGATACTGTGACCTTGCACAAGATTGTCATGCCACAAGCTGCA


TTTGATAACTTTACTGAAGGTACAAAAGGTAAGAATGATAGCGATTATGTTGGTAAACAAATTAATGACCTT


AAATCTTATTTTGGCTCAACCGATGCTAAAGAAATCAAGGGTGCTTTCTTTGTTTTCAAAAATGAAACTGGT


ACAAAATTCATTACTGAAAATGGTAAGGAAGTCGATACTTTGGAAGCTAAAGATGCTGAAGGTGGTGCTGTT


CTTTCAGGGTTAACAAAAGACAATGGTTTTGTTTTTAACACTGCTAAGTTAAAAGGAATTTACCAAATCGTT


GAATTGAAAGAAAAATCAAACTACGATAACAACGGTTCTATCTTGGCTGATTCAAAAGCAGTTCCAGTTAAA


ATCACTCTGCCATTGGTAAACAACCAAGGTGTTGTTAAAGATGCTCACATTTATCCAAAGAATACTGAAACA


AAACCACAAGTAGATAAGAACTTTGCAGATAAAGATCTTGATTATACTGACAACCGAAAAGACAAAGGTGTT


GTCTCAGCGACAGTTGGTGACAAAAAAGAATACATAGTTGGAACAAAAATTCTTAAAGGCTCAGACTATAAG


AAACTGGTTTGGACTGATAGCATGACTAAAGGTTTGACGTTCAACAACAACGTTAAAGTAACATTGGATGGT


GAAGATTTTCCTGTTTTAAACTACAAACTCGTAACAGATGACCAAGGTTTCCGTCTTGCCTTGAATGCAACA


GGTCTTGCAGCAGTAGCAGCAGCTGCAAAAGACAAAGATGTTGAAATCAAGATCACTTACTCAGCTACGGTG


AACGGCTCCACTACTGTTGAAATTCCAGAAACCAATGATGTTAAATTGGACTATGGTAATAACCCAACGGAA


GAAAGTGAACCACAAGAAGGTACTCCAGCTAACCAAGAAATTAAAGTCATTAAAGACTGGGCAGTAGATGGT


ACAATTACTGATGCTAATGTTGCAGTTAAAGCTATCTTTACCTTGCAAGAAAAACAAACGGATGGTACATGG


GTGAACGTTGCTTCACACGAAGCAACAAAACCATCACGCTTTGAACATACTTTCACAGGTTTGGATAATGCT


AAAACTTACCGCGTTGTCGAACGTGTTAGCGGCTACACTCCAGAATACGTATCATTTAAAAATGGTGTTGTG


ACTATCAAGAACAACAAAAACTCAAATGATCCAACTCCAATCAACCCATCAGAACCAAAAGTGGTGACTTAT


GGACGTAAATTTGTGAAAACAAATCAAGCTAACACTGAACGCTTGGCAGGAGCTACCTTCCTCGTTAAGAAA


GAAGGCAAATACTTGGCACGTAAAGCAGGTGCAGCAACTGCTGAAGCAAAGGCAGCTGTAAAAACTGCTAAA


CTAGCATTGGATGAAGCTGTTAAAGCTTATAACGACTTGACTAAAGAAAAACAAGAAGGCCAAGAAGGTAAA


ACAGCATTGGCTACTGTTGATCAAAAACAAAAAGCTTACAATGACGCTTTTGTTAAAGCTAACTACTCATAT


GAATGGGTTGCAGATAAAAAGGCTGATAATGTTGTTAAATTGATCTCTAACGCCGGTGGTCAATTTGAAATT


ACTGGTTTGGATAAAGGCACTTATGGCTTGGAAGAAACTCAAGCACCAGCAGGTTATGCGACATTGTCAGGT


GATGTAAACTTTGAAGTAACTGCCACATCATATAGCAAAGGGGCTACAACTGACATCGCATATGATAAAGGC


TCTGTAAAAAAAGATGCCCAACAAGTTCAAAACAAAAAAGTAACCATCCCACAAACAGGTGGTATTGGTACA


ATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCAGTAGTTATCATGAAAAAACGTCAATCAGAG


GAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 122):


MKKINKCLTMFSTLLLILTSLFSVAPAFADDATTDTVTLHKIVMPQAAFDNFTEGTKGKNDSDYVGKQINDL


KSYFGSTDAKEIKGAFFVFKNETGTKFITENGKEVDTLEAKDAEGGAVLSGLTKDNGFVFNTAKLKGIYQIV


ELKEKSNYDNNGSILADSKAVPVKITLPLVNNQGVVKDAHIYPKNTETKPQVDKNFADKDLDYTDNRKDKGV


VSATVGDKKEYIVGTKILKGSDYKKLVWTDSMTKGLTFNNNVKVTLDGEDFPVLNYKLVTDDQGFRLALNAT


GLAAVAAAAKDKDVEIKITYSATVNGSTTVEIPETNDVKLDYGNNPTEESEPQEGTPANQEIKVIKDWAVDG


TITDANVAVKAIFTLQEKQTDGTWVNVASHEATKPSRFEHTFTGLDNAKTYRVVERVSGYTPEYVSFKNGVV


TIKNNKNSNDPTPINPSEPKVVTYGRKFVKTNQANTERLAGATFLVKKEGKYLARKAGAATAEAKAAVKTAK


LALDEAVKAYNDLTKEKQEGQEGKTALATVDQKQKAYNDAFVKANYSYEWVADKKADNVVKLISNAGGQFEI


TGLDKGTYGLEETQAPAGYATLSGDVNFEVTATSYSKGATTDIAYDKGSVKKDAQQVQNKKVTIPQTGGIGT


ILFTIIGLSIMLGAVVIMKKRQSEEA


 


Strain IC379


ORF DNA sequence (SEQ ID NO: 165):


ATGAAAAAAATCAACAAATGTCTTACAATGTTCTCGACACTGCTATTGATCTTAACGTCACTATTCTCAGTT


GCACCAGCGTTTGCGGACGACGCAACAACTGATACTGTGACCTTGCACAAGATTGTCATGCCACAAGCTGCA


TTTGATAACTTTACTGAAGGTACAAAAGGTAAGAATGATAGCGATTATGTTGGTAAACAAATTAATGACCTT


AAATCTTATTTTGGCTCAACCGATGCTAAAGAAATCAAGGGTGCTTTCTTTGTTTTCAAAAATGAAACTGGT


ACAAAATTCATTACTGAAAATGGTAAGGAAGTCGATACTTTGGAAGCTAAAGATGCTGAAGGTGGTGCTGTT


CTTTCAGGGTTAACAAAAGACAATGGTTTTGTTTTTAACACTGCTAAGTTAAAAGGAATTTACCAAATCGTT


GAATTGAAAGAAAAATCAAACTACGATAACAACGGTTCTATCTTGGCTGATTCAAAAGCAGTTCCAGTTAAA


ATCACTCTGCCATTGGTAAACAACCAAGGTGTTGTTAAAGATGCTCACATTTATCCAAAGAATACTGAAACA


AAACCACAAGTAGATAAGAACTTTGCAGATAAAGATCTTGATTATACTGACAACCGAAAAGACAAAGGTGTT


GTCTCAGCGACAGTTGGTGACAAAAAAGAATACATAGTTGGAACAAAAATTCTTAAAGGCTCAGACTATAAG


AAACTGGTTTGGACTGATAGCATGACTAAAGGTTTGACGTTCAACAACAACGTTAAAGTAACATTGGATGGT


GAAGATTTTCCTGTTTTAAACTACAAACTCGTAACAGATGACCAAGGTTTCCGTCTTGCCTTGAATGCAACA


GGTCTTGCAGCAGTAGCAGCAGCTGCAAAAGACAAAGATGTTGAAATCAAGATCACTTACTCAGCTACGGTG


AACGGCTCCACTACTGTTGAAATTCCAGAAACCAATGATGTTAAATTGGACTATGGTAATAACCCAACGGAA


GAAAGTGAACCACAAGAAGGTACTCCAGCTAACCAAGAAATTAAAGTCATTAAAGACTGGGCAGTAGATGGT


ACAATTACTGATGCTAATGTTGCAGTTAAAGCTATCTTTACCTTGCAAGAAAAACAAACGGATGGTACATGG


GTGAACGTTGCTTCACACGAAGCAACAAAACCATCACGCTTTGAACATACTTTCACAGGTTTGGATAATGCT


AAAACTTACCGCGTTGTCGAACGTGTTAGCGGCTACACTCCAGAATACGTATCATTTAAAAATGGTGTTGTG


ACTATCAAGAACAACAAAAACTCAAATGATCCAACTCCAATCAACCCATCAGAACCAAAAGTGGTGACTTAT


GGACGTAAATTTGTGAAAACAAATCAAGCTAACACTGAACGCTTGGCAGGAGCTACCTTCCTCGTTAAGAAA


GAAGGCAAATACTTGGCACGTAAAGCAGGTGCAGCAACTGCTGAAGCAAAGGCAGCTGTAAAAACTGCTAAA


CTAGCATTGGATGAAGCTGTTAAAGCTTATAACGACTTGACTAAAGAAAAACAAGAAGGCCAAGAAGGTAAA


ACAGCATTGGCTACTGTTGATCAAAAACAAAAAGCTTACAATGACGCTTTTGTTAAAGCTAACTACTCATAT


GAATGGGTTGCAGATAAAAAGGCTGATAATGTTGTTAAATTGATCTCTAACGCCGGTGGTCAATTTGAAATT


ACTGGTTTGGATAAAGGCACTTATGGCTTGGAAGAAACTCAAGCACCAGCAGGTTATGCGACATTGTCAGGT


GATGTAAACTTTGAAGTAACTGCCACATCATATAGCAAAGGGGCTACAACTGACATCGCATATGATAAAGGC


TCTGTAAAAAAAGATGCCCAACAAGTTCAAAACAAAAAAGTAACCATCCCACAAACAGGTGGTATTGGTACA


ATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCAGTAGTTATCATGAAAAAACGTCAATCAGAG


GAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 123):


MKKINKCLTMFSTLLLILTSLFSVAPAFADDATTDTVTLHKIVMPQAAFDNFTEGTKGKNDSDYVGKQINDL


KSYFGSTDAKEIKGAFFVFKNETGTKFITENGKEVDTLEAKDAEGGAVLSGLTKDNGFVFNTAKLKGIYQIV


ELKEKSNYDNNGSILADSKAVPVKITLPLVNNQGVVKDAHIYPKNTETKPQVDKNFADKDLDYTDNRKDKGV


VSATVGDKKEYIVGTKILKGSDYKKLVWTDSMTKGLTFNNNVKVTLDGEDFPVLNYKLVTDDQGFRLALNAT


GLAAVAAAAKDKDVEIKITYSATVNGSTTVEIPETNDVKLDYGNNPTEESEPQEGTPANQEIKVIKDWAVDG


TITDANVAVKAIFTLQEKQTDGTWVNVASHEATKPSRFEHTFTGLDNAKTYRVVERVSGYTPEYVSFKNGVV


TIKNNKNSNDPTPINPSEPKVVTYGRKFVKTNQANTERLAGATFLVKKEGKYLARKAGAATAEAKAAVKTAK


LALDEAVKAYNDLTKEKQEGQEGKTALATVDQKQKAYNDAFVKANYSYEWVADKKADNVVKLISNAGGQFEI


TGLDKGTYGLEETQAPAGYATLSGDVNFEVTATSYSKGATTDIAYDKGSVKKDAQQVQNKKVTIPQTGGIGT


ILFTIIGLSIMLGAVVIMKKRQSEEA


 


Strain IC432


ORF DNA sequence (SEQ ID NO: 166):


ATGAAAAAAATCAACAAATGTCTTACAATGTTCTCGACACTGCTATTGATCTTAACGTCACTATTCTCAGTT


GCACCAGCGTTTGCGGACGACGCAACAACTGATACTGTGACCTTGCACAAGATTGTCATGCCACAAGCTGCA


TTTGATAACTTTACTGAAGGTACAAAAGGTAAGAATGATAGCGATTATGTTGGTAAACAAATTAATGACCTT


AAATCTTATTTTGGCTCAACCGATGCTAAAGAAATCAAGGGTGCTTTCTTTGTTTTCAAAAATGAAACTGGT


ACAAAATTCATTACTGAAAATGGTAAGGAAGTCGATACTTTGGAAGCTAAAGATGCTGAAGGTGGTGCTGTT


CTTTCAGGGTTAACAAAAGACAATGGTTTTGTTTTTAACACTGCTAAGTTAAAAGGAATTTACCAAATCGTT


GAATTGAAAGAAAAATCAAACTACGATAACAACGGTTCTATCTTGGCTGATTCAAAAGCAGTTCCAGTTAAA


ATCACTCTGCCATTGGTAAACAACCAAGGTGTTGTTAAAGATGCTCACATTTATCCAAAGAATACTGAAACA


AAACCACAAGTAGATAAGAACTTTGCAGATAAAGATCTTGATTATACTGACAACCGAAAAGACAAAGGTGTT


GTCTCAGCGACAGTTGGTGACAAAAAAGAATACATAGTTGGAACAAAAATTCTTAAAGGCTCAGACTATAAG


AAACTGGTTTGGACTGATAGCATGACTAAAGGTTTGACGTTCAACAACAACGTTAAAGTAACATTGGATGGT


GAAGATTTTCCTGTTTTAAACTACAAACTCGTAACAGATGACCAAGGTTTCCGTCTTGCCTTGAATGCAACA


GGTCTTGCAGCAGTAGCAGCAGCTGCAAAAGACAAAGATGTTGAAATCAAGATCACTTACTCAGCTACGGTG


AACGGCTCCACTACTGTTGAAATTCCAGAAACCAATGATGTTAAATTGGACTATGGTAATAACCCAACGGAA


GAAAGTGAACCACAAGAAGGTACTCCAGCTAACCAAGAAATTAAAGTCATTAAAGACTGGGCAGTAGATGGT


ACAATTACTGATGCTAATGTTGCAGTTAAAGCTATCTTTACCTTGCAAGAAAAACAAACGGATGGTACATGG


GTGAACGTTGCTTCACACGAAGCAACAAAACCATCACGCTTTGAACATACTTTCACAGGTTTGGATAATGCT


AAAACTTACCGCGTTGTCGAACGTGTTAGCGGCTACACTCCAGAATACGTATCATTTAAAAATGGTGTTGTG


ACTATCAAGAACAACAAAAACTCAAATGATCCAACTCCAATCAACCCATCAGAACCAAAAGTGGTGACTTAT


GGACGTAAATTTGTGAAAACAAATCAAGCTAACACTGAACGCTTGGCAGGAGCTACCTTCCTCGTTAAGAAA


GAAGGCAAATACTTGGCACGTAAAGCAGGTGCAGCAACTGCTGAAGCAAAGGCAGCTGTAAAAACTGCTAAA


CTAGCATTGGATGAAGCTGTTAAAGCTTATAACGACTTGACTAAAGAAAAACAAGAAGGCCAAGAAGGTAAA


ACAGCATTGGCTACTGTTGATCAAAAACAAAAAGCTTACAATGACGCTTTTGTTAAAGCTAACTACTCATAT


GAATGGGTTGCAGATAAAAAGGCTGATAATGTTGTTAAATTGATCTCTAACGCCGGTGGTCAATTTGAAATT


ACTGGTTTGGATAAAGGCACTTATGGCTTGGAAGAAACTCAAGCACCAGCAGGTTATGCGACATTGTCAGGT


GATGTAAACTTTGAAGTAACTGCCACATCATATAGCAAAGGGGCTACAACTGACATCGCATATGATAAAGGC


TCTGTAAAAAAAGATGCCCAACAAGTTCAAAACAAAAAAGTAACCATCCCACAAACAGGTGGTATTGGTACA


ATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCAGTAGTTATCATGAAAAAACGTCAATCAGAG


GAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 124):


MKKINKCLTMFSTLLLILTSLFSVAPAFADDATTDTVTLHKIVMPQAAFDNFTEGTKGKNDSDYVGKQINDL


KSYFGSTDAKEIKGAFFVFKNETGTKFITENGKEVDTLEAKDAEGGAVLSGLTKDNGFVFNTAKLKGIYQIV


ELKEKSNYDNNGSILADSKAVPVKITLPLVNNQGVVKDAHIYPKNTETKPQVDKNFADKDLDYTDNRKDKGV


VSATVGDKKEYIVGTKILKGSDYKKLVWTDSMTKGLTFNNNVKVTLDGEDFPVLNYKLVTDDQGFRLALNAT


GLAAVAAAAKDKDVEIKITYSATVNGSTTVEIPETNDVKLDYGNNPTEESEPQEGTPANQEIKVIKDWAVDG


TITDANVAVKAIFTLQEKQTDGTWVNVASHEATKPSRFEHTFTGLDNAKTYRVVERVSGYTPEYVSFKNGVV


TIKNNKNSNDPTPINPSEPKVVTYGRKFVKTNQANTERLAGATFLVKKEGKYLARKAGAATAEAKAAVKTAK


LALDEAVKAYNDLTKEKQEGQEGKTALATVDQKQKAYNDAFVKANYSYEWVADKKADNVVKLISNAGGQFEI


TGLDKGTYGLEETQAPAGYATLSGDVNFEVTATSYSKGATTDIAYDKGSVKKDAQQVQNKKVTIPQTGGIGT


ILFTIIGLSIMLGAVVIMKKRQSEEA


 


Strain IC455


ORF DNA sequence (SEQ ID NO: 167):


ATGAAAAGAATCAACAAATATTTTGCAATGTTCTCGGCATTGTTACTGACTTTAACGTCATTGCTCTCAGTT


GCACCAGCGTTTGCGGATGAAGCAACAACTAATACAGTGACTTTGCACAAGATTTTGCAAACCGAATCAAAT


CTTAACAAAAGTAACTTCCCAGGAACTACAGGTCTTAACGGAAAAGACTACAAAGGTGGAGCTATTTCTGAC


CTTGCTGGTTACTTTGGCGAGGGATCTAAAGAAATCGAAGGTGCGTTCTTTGCTTTAGCTTTGAAAGAAGAT


AAAAGTGGTAAAGTGCAATATGTTAAGGCAAAAGAAGGTAACAAATTAACACCAGCCTTAATTAATAAAGAT


GGTACTCCTGAAATAACAGTAAATATTGATGAGGCCGTGTCTGGATTGACACCAGAGGGAGATACTGGACTT


GTTTTCAACACCAAAGGATTGAAAGGCGAGTTTAAAATTGTTGAAGTTAAATCAAAATCTACTTACAACAAT


AATGGTTCCCTCCTGGCTGCTTCAAAAGCGGTTCCAGTTAACATCACTCTTCCATTGGTAAATGAAGATGGT


GTTGTTGCTGATGCCCATGTTTATCCAAAGAACACTGAAGAAAAACCAGAAATTGATAAAAACTTTGCTAAA


ACAAACGATTTGACAGCATTGACAGATGTTAATAGACTTTTGACAGCTGGCGCAAATTATGGTAATTATGCA


CGTGACAAAGCAACTGCTACTGCTGAAATCGGTAAAGTTGTTCCTTATGAAGTTAAAACAAAAATTCACAAA


GGTTCTAAATACGAAAACTTGGTTTGGACAGATATAATGTCAAATGGTTTGACAATGGGTTCAACTGTTAGC


CTTAAAGCTTCAGGAACTACAGAAACTTTTGCTAAGGATACAGACTATGAACTTAGCATTGATGCCCGTGGT


TTCACATTAAAATTCACAGCTGATGGATTGGGCAAATTGGAAAAAGCAGCTAAAACAGCTGATATTGAATTT


ACATTGACTTATAGTGCTACTGTTAATGGTCAAGCAATTATTGATAATCCAGAATCCAATGATATCAAATTG


TCGTATGGTAACAAACCAGGTAAAGACTTGACTGAACTTCCTGTTACACCTTCAAAGGGTGAAGTAACAGTT


GCTAAAACTTGGTCTGACGGAATTGCACCTGATGGTGTAAACGTTGTTTACACATTGAAAGATAAAGATAAA


ACTGTTGCTTCAGTATCATTGACAAAAACATCTAAAGGTACAATCGACCTTGGAAATGGTATCAAATTTGAA


GTATCTGGTAACTTCTCGGGTAAATTCACTGGTCTAGAAAACAAATCATACATGATCTCAGAACGTGTTTCT


GGTTACGGAAGTGCAATAAATCTAGAAAATGGTAAAGTAACCATTACCAATACCAAAGATTCTGATAACCCA


ACACCATTGAACCCAACTGAACCAAAAGTTGAAACTCATGGTAAGAAATTTGTCAAAACTAATGAACAAGGT


GACCGTTTGGCTGGTGCACAATTCGTTGTGAAAAACTCAGCAGGTAAATACCTTGCTCTTAAAGCAGATCAA


TCAGAAGGTCAAAAAACTTTAGCTGCTAAGAAAATAGCTTTAGATGAAGCTATCGCTGCTTATAACAAGTTG


TCTGCAACAGACCAAAAAGGTGAAAAAGGAATTACTGCAAAAGAACTTATCAAAACTAAACAAGCAGATTAC


GATGCAGCCTTCATTGAGGCTCGTACAGCTTATGAGTGGATAACAGATAAGGCTAGAGCCATTACCTACACT


TCAAACGATCAAGGTCAATTTGAAGTTACAGGTCTTGCAGACGGTACTTACAACCTTGAAGAAACACTTGCT


CCAGCAGGATTTGCTAAGTTGGCAGGTAATATTAAGTTTGTAGTTAATCAAGGGTCATACATAACAGGTGGT


AACATTGACTACGTTGCTAACAGCAACCAAAAAGATGCGACACGTGTAGAAAATAAAAAGGTAACAATCCCA


CAAACAGGTGGTATTGGTACAATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCAGTAGTTATC


ATGAAAAGACGCCAATCAAAGGAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 125):


MKRINKYFAMFSALLLTLTSLLSVAPAFADEATTNTVTLHKILQTESNLNKSNFPGTTGLNGKDYKGGAISD


LAGYFGEGSKEIEGAFFALALKEDKSGKVQYVKAKEGNKLTPALINKDGTPEITVNIDEAVSGLTPEGDTGL


VFNTKGLKGEFKIVEVKSKSTYNNNGSLLAASKAVPVNITLPLVNEDGVVADAHVYPKNTEEKPEIDKNFAK


TNDLTALTDVNRLLTAGANYGNYARDKATATAEIGKVVPYEVKTKIHKGSKYENLVWTDIMSNGLTMGSTVS


LKASGTTETFAKDTDYELSIDARGFTLKFTADGLGKLEKAAKTADIEFTLTYSATVNGQAIIDNPESNDIKL


SYGNKPGKDLTELPVTPSKGEVTVAKTWSDGIAPDGVNVVYTLKDKDKTVASVSLTKTSKGTIDLGNGIKFE


VSGNFSGKFTGLENKSYMISERVSGYGSAINLENGKVTITNTKDSDNPTPLNPTEPKVETHGKKFVKTNEQG


DRLAGAQFVVKNSAGKYLALKADQSEGQKTLAAKKIALDEAIAAYNKLSATDQKGEKGITAKELIKTKQADY


DAAFIEARTAYEWITDKARAITYTSNDQGQFEVTGLADGTYNLEETLAPAGFAKLAGNIKFVVNQGSYITGG


NIDYVANSNQKDATRVENKKVTIPQTGGIGTILFTIIGLSIMLGAVVIMKRRQSKEA


 


Strain IC457


ORF DNA sequence (SEQ ID NO: 168):


ATGAAAAAAATCAACAAATGTCTTACAATGTTCTCGACACTGCTATTGATCTTAACGTCACTATTCTCAGTT


GCACCAGCGTTTGCGGACGACGCAACAACTGATACTGTGACCTTGCACAAGATTGTCATGCCACAAGCTGCA


TTTGATAACTTTACTGAAGGTACAAAAGGTAAGAATGATAGCGATTATGTTGGTAAACAAATTAATGACCTT


AAATCTTATTTTGGCTCAACCGATGCTAAAGAAATCAAGGGTGCTTTCTTTGTTTTCAAAAATGAAACTGGT


ACAAAATTCATTACTGAAAATGGTAAGGAAGTCGATACTTTGGAAGCTAAAGATGCTGAAGGTGGTGCTGTT


CTTTCAGGGTTAACAAAAGACAATGGTTTTGTTTTTAACACTGCTAAGTTAAAAGGAATTTACCAAATCGTT


GAATTGAAAGAAAAATCAAACTACGATAACAACGGTTCTATCTTGGCTGATTCAAAAGCAGTTCCAGTTAAA


ATCACTCTGCCATTGGTAAACAACCAAGGTGTTGTTAAAGATGCTCACATTTATCCAAAGAATACTGAAACA


AAACCACAAGTAGATAAGAACTTTGCAGATAAAGATCTTGATTATACTGACAACCGAAAAGACAAAGGTGTT


GTCTCAGCGACAGTTGGTGACAAAAAAGAATACATAGTTGGAACAAAAATTCTTAAAGGCTCAGACTATAAG


AAACTGGTTTGGACTGATAGCATGACTAAAGGTTTGACGTTCAACAACAACGTTAAAGTAACATTGGATGGT


GAAGATTTTCCTGTTTTAAACTACAAACTCGTAACAGATGACCAAGGTTTCCGTCTTGCCTTGAATGCAACA


GGTCTTGCAGCAGTAGCAGCAGCTGCAAAAGACAAAGATGTTGAAATCAAGATCACTTACTCAGCTACGGTG


AACGGCTCCACTACTGTTGAAATTCCAGAAACCAATGATGTTAAATTGGACTATGGTAATAACCCAACGGAA


GAAAGTGAACCACAAGAAGGTACTCCAGCTAACCAAGAAATTAAAGTCATTAAAGACTGGGCAGTAGATGGT


ACAATTACTGATGCTAATGTTGCAGTTAAAGCTATCTTTACCTTGCAAGAAAAACAAACGGATGGTACATGG


GTGAACGTTGCTTCACACGAAGCAACAAAACCATCACGCTTTGAACATACTTTCACAGGTTTGGATAATGCT


AAAACTTACCGCGTTGTCGAACGTGTTAGCGGCTACACTCCAGAATACGTATCATTTAAAAATGGTGTTGTG


ACTATCAAGAACAACAAAAACTCAAATGATCCAACTCCAATCAACCCATCAGAACCAAAAGTGGTGACTTAT


GGACGTAAATTTGTGAAAACAAATCAAGCTAACACTGAACGCTTGGCAGGAGCTACCTTCCTCGTTAAGAAA


GAAGGCAAATACTTGGCACGTAAAGCAGGTGCAGCAACTGCTGAAGCAAAGGCAGCTGTAAAAACTGCTAAA


CTAGCATTGGATGAAGCTGTTAAAGCTTATAACGACTTGACTAAAGAAAAACAAGAAGGCCAAGAAGGTAAA


ACAGCATTGGCTACTGTTGATCAAAAACAAAAAGCTTACAATGACGCTTTTGTTAAAGCTAACTACTCATAT


GAATGGGTTGCAGATAAAAAGGCTGATAATGTTGTTAAATTGATCTCTAACGCCGGTGGTCAATTTGAAATT


ACTGGTTTGGATAAAGGCACTTATGGCTTGGAAGAAACTCAAGCACCAGCAGGTTATGCGACATTGTCAGGT


GATGTAAACTTTGAAGTAACTGCCACATCATATAGCAAAGGGGCTACAACTGACATCGCATATGATAAAGGC


TCTGTAAAAAAAGATGCCCAACAAGTTCAAAACAAAAAAGTAACCATCCCACAAACAGGTGGTATTGGTACA


ATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCAGTAGTTATCATGAAAAAACGTCAATCAGAG


GAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 126):


MKKINKCLTMFSTLLLILTSLFSVAPAFADDATTDTVTLHKIVMPQAAFDNFTEGTKGKNDSDYVGKQINDL


KSYFGSTDAKEIKGAFFVFKNETGTKFITENGKEVDTLEAKDAEGGAVLSGLTKDNGFVFNTAKLKGIYQIV


ELKEKSNYDNNGSILADSKAVPVKITLPLVNNQGVVKDAHIYPKNTETKPQVDKNFADKDLDYTDNRKDKGV


VSATVGDKKEYIVGTKILKGSDYKKLVWTDSMTKGLTFNNNVKVTLDGEDFPVLNYKLVTDDQGFRLALNAT


GLAAVAAAAKDKDVEIKITYSATVNGSTTVEIPETNDVKLDYGNNPTEESEPQEGTPANQEIKVIKDWAVDG


TITDANVAVKAIFTLQEKQTDGTWVNVASHEATKPSRFEHTFTGLDNAKTYRVVERVSGYTPEYVSFKNGVV


TIKNNKNSNDPTPINPSEPKVVTYGRKFVKTNQANTERLAGATFLVKKEGKYLARKAGAATAEAKAAVKTAK


LALDEAVKAYNDLTKEKQEGQEGKTALATVDQKQKAYNDAFVKANYSYEWVADKKADNVVKLISNAGGQFEI


TGLDKGTYGLEETQAPAGYATLSGDVNFEVTATSYSKGATTDIAYDKGSVKKDAQQVQNKKVTIPQTGGIGT


ILFTIIGLSIMLGAVVIMKKRQSEEA


 


Strain IC458


ORF DNA sequence (SEQ ID NO: 169):


ATGAAAAAAATCAACAAATATTTTGCAGTCTTCTCGGCCTTGCTACTGACCGTAACATCATTGCTCTCAGTT


GCACCAGCGTTTGCGGACGAAGCAACAACTAATACAGTGACTTTGCACAAGATCTTGCAAACTGAATCAAAT


CTTAATAAAAGTAACTTCCCAGGAACTACAGGCCTTAACGGAGATGACTATAAAGGTGAATCTATTTCTGAC


CTTGCTGAATACTTTGGATCAGGTTCTAAAGAAATTGACGGTGCTTTCTTTGCTTTGGCTTTAGAAGAGGAA


AAAGATGGTGTCGTACAATATGTTAAGGCAAAAGCAAATGACAAATTAACACCAGACTTAATTACTAAAGGT


ACACCTGCAACAACAACAAAAGTTGAAGAAGCTGTAGGTGGTTTGACAACTGGTACGGGTATTGTTTTCAAT


ACAGCTGGTTTGAAAGGTAATTTCAAAATTATTGAATTGAAAGACAAATCAACTTACAACAATAATGGTTCC


CTCTTAGCAGCTTCAAAAGCAGTTCCGGTGAAAATCACTCTTCCATTGGTAAGCAAAGATGGTGTTGTTAAA


GATGCACACGTTTATCCAAAGAACACTGAAACAAAACCAGAAGTAGACAAGAACTTCGCTAAAACAAACGAT


TTGACAGCTCTCAAAGACGCTACTCTTCTTAAGGCTGGTGCAGACTACAAAAACTATTCAGCGACTAAAGCT


ACTGTAACAGCTGAAATCGGTAAAGTTATCCCTTACGAAGTTAAAACAAAAGTTCTTAAAGGTTCTAAATAC


GAAAAACTGGTTTGGACCGATACCATGTCAAATGGTTTGACAATGGGTGATGATGTTAACCTTGCAGTTTCA


GGGACTACAACAACTTTCATTAAAGATATAGATTACACTCTTAGCATTGATGACCGTGGTTTCACATTGAAA


TTCAAAGCTACTGGATTGGACAAATTGGAAGAAGCAGCTAAAGCATCTGATGTTGAATTTACATTGACTTAT


AAAGCTACTGTTAATGGCCAAGCAATTATTGACAACCCAGAAGTCAATGACATCAAATTGGACTATGGTAAT


AAACCTGGTACAGATTTATCAGAACAACCTGTGACACCTGAAGATGGTGAAGTTAAAGTCACTAAAACATGG


GCAGCAGGTGCTAATAAAGCAGACGCTAAAGTTGTCTACACACTTAAAAATGCTACTAAACAAGTCGTAGCT


TCTGTCGCATTGACCGCAGCTGATACAAAAGGTACGATTAATCTTGGTAAAGGCATGACCTTTGAAATCACA


GGAGCTTTCTCAGGTACATTCAAAGGCCTTCAAAATAAAGCTTACACTGTTTCTGAACGTGTTGCAGGTTAT


ACTAATGCTATTAATGTTACTGGTAATGCTGTTGCTATCACCAATACACCAGACAGTGACAATCCAACGCCA


CTTAACCCAACTCAACCAAAAGTTGAAACACATGGTAAGAAATTTGTCAAAGTTGGCGATGCAGATGCCCGC


TTAGCTGGTGCACAATTCGTTGTGAAAAATTCAGCTGGTAAATTCCTTGCTCTTAAAGAAGATGCAGCTGTA


TCAGGAGCTCAAACTGAATTGGCAACTGCTAAAACAGACTTGGATAATGCCATCAAAGCTTACAACGGTTTG


ACAAAAGCGCAGCAAGAAGGTGCTGATGGTACATCAGCAAAAGAACTTATCAACACTAAACAGTCAGCTTAC


GACGCAGCCTTCATCAAAGCACGTACAGCTTATACATGGGTAGATGAAAAAACTAAAGCTATTACCTTCACT


TCAAATAATCAAGGTCAATTTGAAGTTACTGGTCTTGAAGTAGGTTCTTACAAACTTGAAGAAACTCTTGCA


CCAGCAGGTTATGCTAAATTGTCAGGCGACATTGAGTTTACAGTTGGACACGATTCTTACACAAGTGGTGAC


ATCAAGTACAAGACAGATGATGCTAGCAACAATGCACAAAAAGTTTTCAATAAAAAAGTAACCATCCCACAA


ACAGGTGGTATTGGTACAATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCGGTAGTTATCATG


AAAAGACGTCAATCAGAGGAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 127):


MKKINKYFAVFSALLLTVTSLLSVAPAFADEATTNTVTLHKILQTESNLNKSNFPGTTGLNGDDYKGESISD


LAEYFGSGSKEIDGAFFALALEEEKDGVVQYVKAKANDKLTPDLITKGTPATTTKVEEAVGGLTTGTGIVFN


TAGLKGNFKIIELKDKSTYNNNGSLLAASKAVPVKITLPLVSKDGVVKDAHVYPKNTETKPEVDKNFAKTND


LTALKDATLLKAGADYKNYSATKATVTAEIGKVIPYEVKTKVLKGSKYEKLVWTDTMSNGLTMGDDVNLAVS


GTTTTFIKDIDYTLSIDDRGFTLKFKATGLDKLEEAAKASDVEFTLTYKATVNGQAIIDNPEVNDIKLDYGN


KPGTDLSEQPVTPEDGEVKVTKTWAAGANKADAKVVYTLKNATKQVVASVALTAADTKGTINLGKGMTFEIT


GAFSGTFKGLQNKAYTVSERVAGYTNAINVTGNAVAITNTPDSDNPTPLNPTQPKVETHGKKFVKVGDADAR


LAGAQFVVKNSAGKFLALKEDAAVSGAQTELATAKTDLDNAIKAYNGLTKAQQEGADGTSAKELINTKQSAY


DAAFIKARTAYTWVDEKTKAITFTSNNQGQFEVTGLEVGSYKLEETLAPAGYAKLSGDIEFTVGHDSYTSGD


IKYKTDDASNNAQKVFNKKVTIPQTGGIGTILFTIIGLSIMLGAVVIMKRRQSEEA


 


Strain IC459


ORF DNA sequence (SEQ ID NO: 170):


ATGAAAAAAATCAACAAATGTCTTACAATGTTCTCGACACTGCTATTGATCTTAACGTCACTATTCTCAGTT


GCACCAGCGTTTGCGGACGACGCAACAACTGATACTGTGACCTTGCACAAGATTGTCATGCCACAAGCTGCA


TTTGATAACTTTACTGAAGGTACAAAAGGTAAGAATGATAGCGATTATGTTGGTAAACAAATTAATGACCTT


AAATCTTATTTTGGCTCAACCGATGCTAAAGAAATCAAGGGTGCTTTCTTTGTTTTCAAAAATGAAACTGGT


ACAAAATTCATTACTGAAAATGGTAAGGAAGTCGATACTTTGGAAGCTAAAGATGCTGAAGGTGGTGCTGTT


CTTTCAGGGTTAACAAAAGACAATGGTTTTGTTTTTAACACTGCTAAGTTAAAAGGAATTTACCAAATCGTT


GAATTGAAAGAAAAATCAAACTACGATAACAACGGTTCTATCTTGGCTGATTCAAAAGCAGTTCCAGTTAAA


ATCACTCTGCCATTGGTAAACAACCAAGGTGTTGTTAAAGATGCTCACATTTATCCAAAGAATACTGAAACA


AAACCACAAGTAGATAAGAACTTTGCAGATAAAGATCTTGATTATACTGACAACCGAAAAGACAAAGGTGTT


GTCTCAGCGACAGTTGGTGACAAAAAAGAATACATAGTTGGAACAAAAATTCTTAAAGGCTCAGACTATAAG


AAACTGGTTTGGACTGATAGCATGACTAAAGGTTTGACGTTCAACAACAACGTTAAAGTAACATTGGATGGT


GAAGATTTTCCTGTTTTAAACTACAAACTCGTAACAGATGACCAAGGTTTCCGTCTTGCCTTGAATGCAACA


GGTCTTGCAGCAGTAGCAGCAGCTGCAAAAGACAAAGATGTTGAAATCAAGATCACTTACTCAGCTACGGTG


AACGGCTCCACTACTGTTGAAATTCCAGAAACCAATGATGTTAAATTGGACTATGGTAATAACCCAACGGAA


GAAAGTGAACCACAAGAAGGTACTCCAGCTAACCAAGAAATTAAAGTCATTAAAGACTGGGCAGTAGATGGT


ACAATTACTGATGCTAATGTTGCAGTTAAAGCTATCTTTACCTTGCAAGAAAAACAAACGGATGGTACATGG


GTGAACGTTGCTTCACACGAAGCAACAAAACCATCACGCTTTGAACATACTTTCACAGGTTTGGATAATGCT


AAAACTTACCGCGTTGTCGAACGTGTTAGCGGCTACACTCCAGAATACGTATCATTTAAAAATGGTGTTGTG


ACTATCAAGAACAACAAAAACTCAAATGATCCAACTCCAATCAACCCATCAGAACCAAAAGTGGTGACTTAT


GGACGTAAATTTGTGAAAACAAATCAAGCTAACACTGAACGCTTGGCAGGAGCTACCTTCCTCGTTAAGAAA


GAAGGCAAATACTTGGCACGTAAAGCAGGTGCAGCAACTGCTGAAGCAAAGGCAGCTGTAAAAACTGCTAAA


CTAGCATTGGATGAAGCTGTTAAAGCTTATAACGACTTGACTAAAGAAAAACAAGAAGGCCAAGAAGGTAAA


ACAGCATTGGCTACTGTTGATCAAAAACAAAAAGCTTACAATGACGCTTTTGTTAAAGCTAACTACTCATAT


GAATGGGTTGCAGATAAAAAGGCTGATAATGTTGTTAAATTGATCTCTAACGCCGGTGGTCAATTTGAAATT


ACTGGTTTGGATAAAGGCACTTATGGCTTGGAAGAAACTCAAGCACCAGCAGGTTATGCGACATTGTCAGGT


GATGTAAACTTTGAAGTAACTGCCACATCATATAGCAAAGGGGCTACAACTGACATCGCATATGATAAAGGC


TCTGTAAAAAAAGATGCCCAACAAGTTCAAAACAAAAAAGTAACCATCCCACAAACAGGTGGTATTGGTACA


ATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCAGTAGTTATCATGAAAAAACGTCAATCAGAG


GAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 128):


MKKINKCLTMFSTLLLILTSLFSVAPAFADDATTDTVTLHKIVMPQAAFDNFTEGTKGKNDSDYVGKQINDL


KSYFGSTDAKEIKGAFFVFKNETGTKFITENGKEVDTLEAKDAEGGAVLSGLTKDNGFVFNTAKLKGIYQIV


ELKEKSNYDNNGSILADSKAVPVKITLPLVNNQGVVKDAHIYPKNTETKPQVDKNFADKDLDYTDNRKDKGV


VSATVGDKKEYIVGTKILKGSDYKKLVWTDSMTKGLTFNNNVKVTLDGEDFPVLNYKLVTDDQGFRLALNAT


GLAAVAAAAKDKDVEIKITYSATVNGSTTVEIPETNDVKLDYGNNPTEESEPQEGTPANQEIKVIKDWAVDG


TITDANVAVKAIFTLQEKQTDGTWVNVASHEATKPSRFEHTFTGLDNAKTYRVVERVSGYTPEYVSFKNGVV


TIKNNKNSNDPTPINPSEPKVVTYGRKFVKTNQANTERLAGATFLVKKEGKYLARKAGAATAEAKAAVKTAK


LALDEAVKAYNDLTKEKQEGQEGKTALATVDQKQKAYNDAFVKANYSYEWVADKKADNVVKLISNAGGQFEI


TGLDKGTYGLEETQAPAGYATLSGDVNFEVTATSYSKGATTDIAYDKGSVKKDAQQVQNKKVTIPQTGGIGT


ILFTIIGLSIMLGAVVIMKKRQSEEA


 


Strain IC460


ORF DNA sequence (SEQ ID NO: 171):


ATGAAAAGAATCAACAAATATTTTGCAATGTTCTCGGCATTGTTACTGACTTTAACGTCATTGCTCTCAGTT


GCACCAGCGTTTGCGGATGAAGCAACAACTAATACAGTGACTTTGCACAAGATTTTGCAAACCGAATCAAAT


CTTAACAAAAGTAACTTCCCAGGAACTACAGGTCTTAACGGAAAAGACTACAAAGGTGGAGCTATTTCTGAC


CTTGCTGGTTACTTTGGCGAGGGATCTAAAGAAATCGAAGGTGCGTTCTTTGCTTTAGCTTTGAAAGAAGAT


AAAAGTGGTAAAGTGCAATATGTTAAGGCAAAAGAAGGTAACAAATTAACACCAGCCTTAATTAATAAAGAT


GGTACTCCTGAAATAACAGTAAATATTGATGAGGCCGTGTCTGGATTGACACCAGAGGGAGATACTGGACTT


GTTTTCAACACCAAAGGATTGAAAGGCGAGTTTAAAATTGTTGAAGTTAAATCAAAATCTACTTACAACAAT


AATGGTTCCCTCCTGGCTGCTTCAAAAGCGGTTCCAGTTAACATCACTCTTCCATTGGTAAATGAAGATGGT


GTTGTTGCTGATGCCCATGTTTATCCAAAGAACACTGAAGAAAAACCAGAAATTGATAAAAACTTTGCTAAA


ACAAACGATTTGACAGCATTGACAGATGTTAATAGACTTTTGACAGCTGGCGCAAATTATGGTAATTATGCA


CGTGACAAAGCAACTGCTACTGCTGAAATCGGTAAAGTTGTTCCTTATGAAGTTAAAACAAAAATTCACAAA


GGTTCTAAATACGAAAACTTGGTTTGGACAGATATAATGTCAAATGGTTTGACAATGGGTTCAACTGTTAGC


CTTAAAGCTTCAGGAACTACAGAAACTTTTGCTAAGGATACAGACTATGAACTTAGCATTGATGCCCGTGGT


TTCACATTAAAATTCACAGCTGATGGATTGGGCAAATTGGAAAAAGCAGCTAAAACAGCTGATATTGAATTT


ACATTGACTTATAGTGCTACTGTTAATGGTCAAGCAATTATTGATAATCCAGAATCCAATGATATCAAATTG


TCGTATGGTAACAAACCAGGTAAAGACTTGACTGAACTTCCTGTTACACCTTCAAAGGGTGAAGTAACAGTT


GCTAAAACTTGGTCTGACGGAATTGCACCTGATGGTGTAAACGTTGTTTACACATTGAAAGATAAAGATAAA


ACTGTTGCTTCAGTATCATTGACAAAAACATCTAAAGGTACAATCGACCTTGGAAATGGTATCAAATTTGAA


GTATCTGGTAACTTCTCGGGTAAATTCACTGGTCTAGAAAACAAATCATACATGATCTCAGAACGTGTTTCT


GGTTACGGAAGTGCAATAAATCTAGAAAATGGTAAAGTAACCATTACCAATACCAAAGATTCTGATAACCCA


ACACCATTGAACCCAACTGAACCAAAAGTTGAAACTCATGGTAAGAAATTTGTCAAAACTAATGAACAAGGT


GACCGTTTGGCTGGTGCACAATTCGTTGTGAAAAACTCAGCAGGTAAATACCTTGCTCTTAAAGCAGATCAA


TCAGAAGGTCAAAAAACTTTAGCTGCTAAGAAAATAGCTTTAGATGAAGCTATCGCTGCTTATAACAAGTTG


TCTGCAACAGACCAAAAAGGTGAAAAAGGAATTACTGCAAAAGAACTTATCAAAACTAAACAAGCAGATTAC


GATGCAGCCTTCATTGAGGCTCGTACAGCTTATGAGTGGATAACAGATAAGGCTAGAGCCATTACCTACACT


TCAAACGATCAAGGTCAATTTGAAGTTACAGGTCTTGCAGACGGTACTTACAACCTTGAAGAAACACTTGCT


CCAGCAGGATTTGCTAAGTTGGCAGGTAATATTAAGTTTGTAGTTAATCAAGGGTCATACATAACAGGTGGT


AACATTGACTACGTTGCTAACAGCAACCAAAAAGATGCGACACGTGTAGAAAATAAAAAGGTAACAATCCCA


CAAACAGGTGGTATTGGTACAATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCAGTAGTTATC


ATGAAAAGACGCCAATCAAAGGAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 129):


MKRINKYFAMFSALLLTLTSLLSVAPAFADEATTNTVTLHKILQTESNLNKSNFPGTTGLNGKDYKGGAISD


LAGYFGEGSKEIEGAFFALALKEDKSGKVQYVKAKEGNKLTPALINKDGTPEITVNIDEAVSGLTPEGDTGL


VFNTKGLKGEFKIVEVKSKSTYNNNGSLLAASKAVPVNITLPLVNEDGVVADAHVYPKNTEEKPEIDKNFAK


TNDLTALTDVNRLLTAGANYGNYARDKATATAEIGKVVPYEVKTKIHKGSKYENLVWTDIMSNGLTMGSTVS


LKASGTTETFAKDTDYELSIDARGFTLKFTADGLGKLEKAAKTADIEFTLTYSATVNGQAIIDNPESNDIKL


SYGNKPGKDLTELPVTPSKGEVTVAKTWSDGIAPDGVNVVYTLKDKDKTVASVSLTKTSKGTIDLGNGIKFE


VSGNFSGKFTGLENKSYMISERVSGYGSAINLENGKVTITNTKDSDNPTPLNPTEPKVETHGKKFVKTNEQG


DRLAGAQFVVKNSAGKYLALKADQSEGQKTLAAKKIALDEAIAAYNKLSATDQKGEKGITAKELIKTKQADY


DAAFIEARTAYEWITDKARAITYTSNDQGQFEVTGLADGTYNLEETLAPAGFAKLAGNIKFVVNQGSYITGG


NIDYVANSNQKDATRVENKKVTIPQTGGIGTILFTIIGLSIMLGAVVIMKRRQSKEA


 


Strain IC461


ORF DNA sequence (SEQ ID NO: 172):


ATGAAAAAAATCAACAAATGTCTTACAATGTTCTCGACACTGCTATTGATCTTAACGTCACTATTCTCAGTT


GCACCAGCGTTTGCGGACGACGCAACAACTGATACTGTGACCTTGCACAAGATTGTCATGCCACAAGCTGCA


TTTGATAACTTTACTGAAGGTACAAAAGGTAAGAATGATAGCGATTATGTTGGTAAACAAATTAATGACCTT


AAATCTTATTTTGGCTCAACCGATGCTAAAGAAATCAAGGGTGCTTTCTTTGTTTTCAAAAATGAAACTGGT


ACAAAATTCATTACTGAAAATGGTAAGGAAGTCGATACTTTGGAAGCTAAAGATGCTGAAGGTGGTGCTGTT


CTTTCAGGGTTAACAAAAGACAATGGTTTTGTTTTTAACACTGCTAAGTTAAAAGGAATTTACCAAATCGTT


GAATTGAAAGAAAAATCAAACTACGATAACAACGGTTCTATCTTGGCTGATTCAAAAGCAGTTCCAGTTAAA


ATCACTCTGCCATTGGTAAACAACCAAGGTGTTGTTAAAGATGCTCACATTTATCCAAAGAATACTGAAACA


AAACCACAAGTAGATAAGAACTTTGCAGATAAAGATCTTGATTATACTGACAACCGAAAAGACAAAGGTGTT


GTCTCAGCGACAGTTGGTGACAAAAAAGAATACATAGTTGGAACAAAAATTCTTAAAGGCTCAGACTATAAG


AAACTGGTTTGGACTGATAGCATGACTAAAGGTTTGACGTTCAACAACAACGTTAAAGTAACATTGGATGGT


GAAGATTTTCCTGTTTTAAACTACAAACTCGTAACAGATGACCAAGGTTTCCGTCTTGCCTTGAATGCAACA


GGTCTTGCAGCAGTAGCAGCAGCTGCAAAAGACAAAGATGTTGAAATCAAGATCACTTACTCAGCTACGGTG


AACGGCTCCACTACTGTTGAAATTCCAGAAACCAATGATGTTAAATTGGACTATGGTAATAACCCAACGGAA


GAAAGTGAACCACAAGAAGGTACTCCAGCTAACCAAGAAATTAAAGTCATTAAAGACTGGGCAGTAGATGGT


ACAATTACTGATGCTAATGTTGCAGTTAAAGCTATCTTTACCTTGCAAGAAAAACAAACGGATGGTACATGG


GTGAACGTTGCTTCACACGAAGCAACAAAACCATCACGCTTTGAACATACTTTCACAGGTTTGGATAATGCT


AAAACTTACCGCGTTGTCGAACGTGTTAGCGGCTACACTCCAGAATACGTATCATTTAAAAATGGTGTTGTG


ACTATCAAGAACAACAAAAACTCAAATGATCCAACTCCAATCAACCCATCAGAACCAAAAGTGGTGACTTAT


GGACGTAAATTTGTGAAAACAAATCAAGCTAACACTGAACGCTTGGCAGGAGCTACCTTCCTCGTTAAGAAA


GAAGGCAAATACTTGGCACGTAAAGCAGGTGCAGCAACTGCTGAAGCAAAGGCAGCTGTAAAAACTGCTAAA


CTAGCATTGGATGAAGCTGTTAAAGCTTATAACGACTTGACTAAAGAAAAACAAGAAGGCCAAGAAGGTAAA


ACAGCATTGGCTACTGTTGATCAAAAACAAAAAGCTTACAATGACGCTTTTGTTAAAGCTAACTACTCATAT


GAATGGGTTGCAGATAAAAAGGCTGATAATGTTGTTAAATTGATCTCTAACGCCGGTGGTCAATTTGAAATT


ACTGGTTTGGATAAAGGCACTTATGGCTTGGAAGAAACTCAAGCACCAGCAGGTTATGCGACATTGTCAGGT


GATGTAAACTTTGAAGTAACTGCCACATCATATAGCAAAGGGGCTACAACTGACATCGCATATGATAAAGGC


TCTGTAAAAAAAGATGCCCAACAAGTTCAAAACAAAAAAGTAACCATCCCACAAACAGGTGGTATTGGTACA


ATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCAGTAGTTATCATGAAAAAACGTCAATCAGAG


GAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 130):


MKKINKCLTMFSTLLLILTSLFSVAPAFADDATTDTVTLHKIVMPQAAFDNFTEGTKGKNDSDYVGKQINDL


KSYFGSTDAKEIKGAFFVFKNETGTKFITENGKEVDTLEAKDAEGGAVLSGLTKDNGFVFNTAKLKGIYQIV


ELKEKSNYDNNGSILADSKAVPVKITLPLVNNQGVVKDAHIYPKNTETKPQVDKNFADKDLDYTDNRKDKGV


VSATVGDKKEYIVGTKILKGSDYKKLVWTDSMTKGLTFNNNVKVTLDGEDFPVLNYKLVTDDQGFRLALNAT


GLAAVAAAAKDKDVEIKITYSATVNGSTTVEIPETNDVKLDYGNNPTEESEPQEGTPANQEIKVIKDWAVDG


TITDANVAVKAIFTLQEKQTDGTWVNVASHEATKPSRFEHTFTGLDNAKTYRVVERVSGYTPEYVSFKNGVV


TIKNNKNSNDPTPINPSEPKVVTYGRKFVKTNQANTERLAGATFLVKKEGKYLARKAGAATAEAKAAVKTAK


LALDEAVKAYNDLTKEKQEGQEGKTALATVDQKQKAYNDAFVKANYSYEWVADKKADNVVKLISNAGGQFEI


TGLDKGTYGLEETQAPAGYATLSGDVNFEVTATSYSKGATTDIAYDKGSVKKDAQQVQNKKVTIPQTGGIGT


ILFTIIGLSIMLGAVVIMKKRQSEEA


 


Strain IC462


ORF DNA sequence (SEQ ID NO: 173):


ATGAAAAGAATCAACAAATATTTTGCAATGTTCTCGGCATTGTTACTGACTTTAACGTCATTGCTCTCAGTT


GCACCAGCGTTTGCGGATGAAGCAACAACTAATACAGTGACTTTGCACAAGATTTTGCAAACCGAATCAAAT


CTTAACAAAAGTAACTTCCCAGGAACTACAGGTCTTAACGGAAAAGACTACAAAGGTGGAGCTATTTCTGAC


CTTGCTGGTTACTTTGGCGAGGGATCTAAAGAAATCGAAGGTGCGTTCTTTGCTTTAGCTTTGAAAGAAGAT


AAAAGTGGTAAAGTGCAATATGTTAAGGCAAAAGAAGGTAACAAATTAACACCAGCCTTAATTAATAAAGAT


GGTACTCCTGAAATAACAGTAAATATTGATGAGGCCGTGTCTGGATTGACACCAGAGGGAGATACTGGACTT


GTTTTCAACACCAAAGGATTGAAAGGCGAGTTTAAAATTGTTGAAGTTAAATCAAAATCTACTTACAACAAT


AATGGTTCCCTCCTGGCTGCTTCAAAAGCGGTTCCAGTTAACATCACTCTTCCATTGGTAAATGAAGATGGT


GTTGTTGCTGATGCCCATGTTTATCCAAAGAACACTGAAGAAAAACCAGAAATTGATAAAAACTTTGCTAAA


ACAAACGATTTGACAGCATTGACAGATGTTAATAGACTTTTGACAGCTGGCGCAAATTATGGTAATTATGCA


CGTGACAAAGCAACTGCTACTGCTGAAATCGGTAAAGTTGTTCCTTATGAAGTTAAAACAAAAATTCACAAA


GGTTCTAAATACGAAAACTTGGTTTGGACAGATATAATGTCAAATGGTTTGACAATGGGTTCAACTGTTAGC


CTTAAAGCTTCAGGAACTACAGAAACTTTTGCTAAGGATACAGACTATGAACTTAGCATTGATGCCCGTGGT


TTCACATTAAAATTCACAGCTGATGGATTGGGCAAATTGGAAAAAGCAGCTAAAACAGCTGATATTGAATTT


ACATTGACTTATAGTGCTACTGTTAATGGTCAAGCAATTATTGATAATCCAGAATCCAATGATATCAAATTG


TCGTATGGTAACAAACCAGGTAAAGACTTGACTGAACTTCCTGTTACACCTTCAAAGGGTGAAGTAACAGTT


GCTAAAACTTGGTCTGACGGAATTGCACCTGATGGTGTAAACGTTGTTTACACATTGAAAGATAAAGATAAA


ACTGTTGCTTCAGTATCATTGACAAAAACATCTAAAGGTACAATCGACCTTGGAAATGGTATCAAATTTGAA


GTATCTGGTAACTTCTCGGGTAAATTCACTGGTCTAGAAAACAAATCATACATGATCTCAGAACGTGTTTCT


GGTTACGGAAGTGCAATAAATCTAGAAAATGGTAAAGTAACCATTACCAATACCAAAGATTCTGATAACCCA


ACACCATTGAACCCAACTGAACCAAAAGTTGAAACTCATGGTAAGAAATTTGTCAAAACTAATGAACAAGGT


GACCGTTTGGCTGGTGCACAATTCGTTGTGAAAAACTCAGCAGGTAAATACCTTGCTCTTAAAGCAGATCAA


TCAGAAGGTCAAAAAACTTTAGCTGCTAAGAAAATAGCTTTAGATGAAGCTATCGCTGCTTATAACAAGTTG


TCTGCAACAGACCAAAAAGGTGAAAAAGGAATTACTGCAAAAGAACTTATCAAAACTAAACAAGCAGATTAC


GATGCAGCCTTCATTGAGGCTCGTACAGCTTATGAGTGGATAACAGATAAGGCTAGAGCCATTACCTACACT


TCAAACGATCAAGGTCAATTTGAAGTTACAGGTCTTGCAGACGGTACTTACAACCTTGAAGAAACACTTGCT


CCAGCAGGATTTGCTAAGTTGGCAGGTAATATTAAGTTTGTAGTTAATCAAGGGTCATACATAACAGGTGGT


AACATTGACTACGTTGCTAACAGCAACCAAAAAGATGCGACACGTGTAGAAAATAAAAAGGTAACAATCCCA


CAAACAGGTGGTATTGGTACAATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCAGTAGTTATC


ATGAAAAGACGCCAATCAAAGGAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 131):


MKRINKYFAMFSALLLTLTSLLSVAPAFADEATTNTVTLHKILQTESNLNKSNFPGTTGLNGKDYKGGAISD


LAGYFGEGSKEIEGAFFALALKEDKSGKVQYVKAKEGNKLTPALINKDGTPEITVNIDEAVSGLTPEGDTGL


VFNTKGLKGEFKIVEVKSKSTYNNNGSLLAASKAVPVNITLPLVNEDGVVADAHVYPKNTEEKPEIDKNFAK


TNDLTALTDVNRLLTAGANYGNYARDKATATAEIGKVVPYEVKTKIHKGSKYENLVWTDIMSNGLTMGSTVS


LKASGTTETFAKDTDYELSIDARGFTLKFTADGLGKLEKAAKTADIEFTLTYSATVNGQAIIDNPESNDIKL


SYGNKPGKDLTELPVTPSKGEVTVAKTWSDGIAPDGVNVVYTLKDKDKTVASVSLTKTSKGTIDLGNGIKFE


VSGNFSGKFTGLENKSYMISERVSGYGSAINLENGKVTITNTKDSDNPTPLNPTEPKVETHGKKFVKTNEQG


DRLAGAQFVVKNSAGKYLALKADQSEGQKTLAAKKIALDEAIAAYNKLSATDQKGEKGITAKELIKTKQADY


DAAFIEARTAYEWITDKARAITYTSNDQGQFEVTGLADGTYNLEETLAPAGFAKLAGNIKFVVNQGSYITGG


NIDYVANSNQKDATRVENKKVTIPQTGGIGTILFTIIGLSIMLGAVVIMKRRQSKEA


 


Strain IC470


ORF DNA sequence (SEQ ID NO: 174):


ATGAAAAAAATCAACAAATGTCTTACAATGTTCTCGACACTGCTATTGATCTTAACGTCACTATTCTCAGTT


GCACCAGCGTTTGCGGACGACGCAACAACTGATACTGTGACCTTGCACAAGATTGTCATGCCACAAGCTGCA


TTTGATAACTTTACTGAAGGTACAAAAGGTAAGAATGATAGCGATTATGTTGGTAAACAAATTAATGACCTT


AAATCTTATTTTGGCTCAACCGATGCTAAAGAAATCAAGGGTGCTTTCTTTGTTTTCAAAAATGAAACTGGT


ACAAAATTCATTACTGAAAATGGTAAGGAAGTCGATACTTTGGAAGCTAAAGATGCTGAAGGTGGTGCTGTT


CTTTCAGGGTTAACAAAAGACAATGGTTTTGTTTTTAACACTGCTAAGTTAAAAGGAATTTACCAAATCGTT


GAATTGAAAGAAAAATCAAACTACGATAACAACGGTTCTATCTTGGCTGATTCAAAAGCAGTTCCAGTTAAA


ATCACTCTGCCATTGGTAAACAACCAAGGTGTTGTTAAAGATGCTCACATTTATCCAAAGAATACTGAAACA


AAACCACAAGTAGATAAGAACTTTGCAGATAAAGATCTTGATTATACTGACAACCGAAAAGACAAAGGTGTT


GTCTCAGCGACAGTTGGTGACAAAAAAGAATACATAGTTGGAACAAAAATTCTTAAAGGCTCAGACTATAAG


AAACTGGTTTGGACTGATAGCATGACTAAAGGTTTGACGTTCAACAACAACGTTAAAGTAACATTGGATGGT


GAAGATTTTCCTGTTTTAAACTACAAACTCGTAACAGATGACCAAGGTTTCCGTCTTGCCTTGAATGCAACA


GGTCTTGCAGCAGTAGCAGCAGCTGCAAAAGACAAAGATGTTGAAATCAAGATCACTTACTCAGCTACGGTG


AACGGCTCCACTACTGTTGAAATTCCAGAAACCAATGATGTTAAATTGGACTATGGTAATAACCCAACGGAA


GAAAGTGAACCACAAGAAGGTACTCCAGCTAACCAAGAAATTAAAGTCATTAAAGACTGGGCAGTAGATGGT


ACAATTACTGATGCTAATGTTGCAGTTAAAGCTATCTTTACCTTGCAAGAAAAACAAACGGATGGTACATGG


GTGAACGTTGCTTCACACGAAGCAACAAAACCATCACGCTTTGAACATACTTTCACAGGTTTGGATAATGCT


AAAACTTACCGCGTTGTCGAACGTGTTAGCGGCTACACTCCAGAATACGTATCATTTAAAAATGGTGTTGTG


ACTATCAAGAACAACAAAAACTCAAATGATCCAACTCCAATCAACCCATCAGAACCAAAAGTGGTGACTTAT


GGACGTAAATTTGTGAAAACAAATCAAGCTAACACTGAACGCTTGGCAGGAGCTACCTTCCTCGTTAAGAAA


GAAGGCAAATACTTGGCACGTAAAGCAGGTGCAGCAACTGCTGAAGCAAAGGCAGCTGTAAAAACTGCTAAA


CTAGCATTGGATGAAGCTGTTAAAGCTTATAACGACTTGACTAAAGAAAAACAAGAAGGCCAAGAAGGTAAA


ACAGCATTGGCTACTGTTGATCAAAAACAAAAAGCTTACAATGACGCTTTTGTTAAAGCTAACTACTCATAT


GAATGGGTTGCAGATAAAAAGGCTGATAATGTTGTTAAATTGATCTCTAACGCCGGTGGTCAATTTGAAATT


ACTGGTTTGGATAAAGGCACTTATGGCTTGGAAGAAACTCAAGCACCAGCAGGTTATGCGACATTGTCAGGT


GATGTAAACTTTGAAGTAACTGCCACATCATATAGCAAAGGGGCTACAACTGACATCGCATATGATAAAGGC


TCTGTAAAAAAAGATGCCCAACAAGTTCAAAACAAAAAAGTAACCATCCCACAAACAGGTGGTATTGGTACA


ATTCTTTTCACAATTATTGGTTTAAGCATTATGCTTGGAGCAGTAGTTATCATGAAAAAACGTCAATCAGAG


GAAGCTTAA


 


ORF amino acid sequence (SEQ ID NO: 132):


MKKINKCLTMFSTLLLILTSLFSVAPAFADDATTDTVTLHKIVMPQAAFDNFTEGTKGKNDSDYVGKQINDL


KSYFGSTDAKEIKGAFFVFKNETGTKFITENGKEVDTLEAKDAEGGAVLSGLTKDNGFVFNTAKLKGIYQIV


ELKEKSNYDNNGSILADSKAVPVKITLPLVNNQGVVKDAHIYPKNTETKPQVDKNFADKDLDYTDNRKDKGV


VSATVGDKKEYIVGTKILKGSDYKKLVWTDSMTKGLTFNNNVKVTLDGEDFPVLNYKLVTDDQGFRLALNAT


GLAAVAAAAKDKDVEIKITYSATVNGSTTVEIPETNDVKLDYGNNPTEESEPQEGTPANQEIKVIKDWAVDG


TITDANVAVKAIFTLQEKQTDGTWVNVASHEATKPSRFEHTFTGLDNAKTYRVVERVSGYTPEYVSFKNGVV


TIKNNKNSNDPTPINPSEPKVVTYGRKFVKTNQANTERLAGATFLVKKEGKYLARKAGAATAEAKAAVKTAK


LALDEAVKAYNDLTKEKQEGQEGKTALATVDQKQKAYNDAFVKANYSYEWVADKKADNVVKLISNAGGQFEI


TGLDKGTYGLEETQAPAGYATLSGDVNFEVTATSYSKGATTDIAYDKGSVKKDAQQVQNKKVTIPQTGGIGT


ILFTIIGLSIMLGAVVIMKKRQSEEA
















TABLE 11





Amino acid and encoding DNA sequences of gbs1478 proteins derived from


different strains of S. agalactiae







Strain 12401


ORF DNA sequence (SEQ ID NO: 204):


ATGAGAAAATACCAAAAATTTTCTAAAATATTGACGTTAAGTCTTTTTTGTTTGTCGCAAATACCGCTTAAT


ACCAATGTTTTAGGGGAAAGTACCGTACCGGAAAATGGTGCTAAAGGAAAGTTAGTTGTTAAAAAGACAGAT


GACCAGAACAAACCACTTTCAAAAGCTACCTTTGTTTTAAAACCTACTTCACACTCAGAAAGCAAAGTAGAA


AAAGTAACTACTGAGGTAACAGGTGAAGCTACTTTTGATAATCTCACACCTGGAGATTACACTTTATCAGAA


GAAACGGCACCCGAAGGATACAAAAAGACTACCCAGACTTGGCAAGTTAAGGTTGAGAGTAATGGAAAAACT


ACGATACAAAATAGTGATGATAAAAAATCTATAATTGAACAAAGGCAAGAGGAACTAGATAAGCAGTATCCC


CTTACAGGAGCTTATGAAGATACAAAAGAATCTTATAATCTTGAGCATGTTAAAAATTCAATTCCAAATGGG


AAATTAGAGGCAAAAGCAGTTAATCCATATTCAAGTGAAGGTGAGCACATAAGAGAAATTCAAGAGGGAACA


TTATCTAAACGTATTTCAGAAGTAAATGATTTGGATCATAATAAATATAAAATTGAGTTAACTGTTAGCGGT


AAATCCATAATAAAAACTATAAATAAAGATGAACCTCTGGATGTTGTTTTTGTTCTTGATAATTCAAATTCT


ATGAAGAATAATGGAAAAAATAACAAGGCAAAAAAGGCAGGTGAAGCAGTAGAAACAATTATAAAAGATGTT


TTAGGAGCAAATGTTGAAAACCGAGCAGCTTTAGTTACTTATGGTTCAGATATTTTTGATGGAAGGACAGTT


AAAGTTATAAAAGGTTTTAAAGAGGATCCTTATTATGGACTTGAAACTAGTTTCACAGTTCAGACAAATGAT


TATAGCTATAAAAAGTTCACTAATATTGCTGCTGATATTATAAAAAAGATCCCTAAAGAAGCTCCAGAAGCT


AAGTGGGGGGGGACAAGTCTAGGATTAACTCCAGAAAAAAAGAGGGAATATGATTTAAGTAAAGTAGGTGAG


ACCTTTACAATGAAAGCTTTTATGGAGGCAGATACCTTGTTAAGTAGTATACAGCGTAAGAGTAGAAAGATT


ATTGTTCATCTAACTGACGGTGTTCCAACAAGATCATATGCCATTAATAGTTTTGTAAAAGGTTCAACATAC


GCAAATCAATTTGAGAGAATAAAAGAAAAAGGTTATTTAGACAAAAATAATTATTTTATAACTGATGATCCA


GAAAAGATCAAAGGCAATGGGGAGAGTTACTTTTTGTTTCCCTTAGATAGTTATCAAACACAGATAATTTCT


GGAAACTTACAAAAACTTCATTATTTAGATTTAAATCTTAATTACCCTAAAGGTACAATTTATAGAAATGGA


CCAGTAAGAGAACATGGAACACCAACCAAACTTTATATAAATAGTTTAAAACAGAAAAATTATGACATCTTT


AATTTTGGTATAGATATATCTGGTTTTAGACAAGTTTATAATGAGGATTATAAGAAAAATCAAGATGGTACT


TTTCAAAAATTGAAGGAGGAAGCTTTTGAACTTTCAGATGGGGAAATAACAGAACTAATGAATTCATTCTCT


TCTAAACCTGAGTATTATACCCCGATAGTAACTTCAGCTGATGTATCTAATAATGAAATTTTATCTAAAATT


CAGCAACAATTTGAAAAGATTTTAACAAAGGAAAACTCAATTGTTAATGGAACTATAGAAGATCCTATGGGT


GATAAAATCAATTTACATCTTGGCAACGGACAAACATTGCAACCAAGTGATTATACTTTACAGGGAAATGAT


GGAAGTATAATGAAAGATAGCATTGCAACTGGAGGGCCTAATAATGATGGTGGGATACTTAAAGGGGTTAAA


TTAGAATACATCAAAAATAAACTCTACGTTAGAGGTTTGAACTTAGGGGAGGGGCAAAAAGTAACACTCACA


TATGATGTGAAACTAGATGACAGTTTTATTAGTAACAAATTCTATGACACTAATGGTAGAACAACATTGAAT


CCTAAATCAGAGGAACCCGATACACTTAGAGATTTTCCAATCCCTAAAATTCGTGATGTGAGAGAATATCCT


ACAATAACGATTAAAAACGAGAAGAAGTTAGGTGAAATTGAATTTACAAAAGTTGATAAAGATAATAATAAG


TTGCTTCTCAAAGGAGCTACATTTGAACTTCAAGAATTTAATGAAGATTATAAACTTTATTTACCAATAAAA


AATAATAATTCAAAAGTAGTGACGGGAGAAAACGGCAAAATTTCTTACAAAGATTTGAAAGATGGCAAATAT


CAGTTAATAGAAGCAGTTTCGCCGAAGGATTATCAAAAAATTACTAATAAACCAATTTTAACTTTTGAAGTT


GTTAAAGGATCGATACAAAATATAATAGCTGTTAATAAACAGATTTCTGAATATCATGAGGAAGGTGACAAG


CATTTAATTACCAACACGCATATTCCACCAAAAGGAATTATTCCGATGACAGGTGGGAAAGGAATTCTATCT


TTCATTTTAATAGGTGGAGCTATGATGTCTATTGCAGGTGGAATTTATATTTGGAAAAGACATAAGAAATCT


AGTGATGCATCAATCGAGAAAGATTAA


 


ORF amino acid sequence (SEQ ID NO: 185):


MRKYQKFSKILTLSLFCLSQIPLNTNVLGESTVPENGAKGKLVVKKTDDQNKPLSKATFVLKPTSHSESKVE


KVTTEVTGEATFDNLTPGDYTLSEETAPEGYKKTTQTWQVKVESNGKTTIQNSDDKKSIIEQRQEELDKQYP


LTGAYEDTKESYNLEHVKNSIPNGKLEAKAVNPYSSEGEHIREIQEGTLSKRISEVNDLDHNKYKIELTVSG


KSIIKTINKDEPLDVVFVLDNSNSMKNNGKNNKAKKAGEAVETIIKDVLGANVENRAALVTYGSDIFDGRTV


KVIKGFKEDPYYGLETSFTVQTNDYSYKKFTNIAADIIKKIPKEAPEAKWGGTSLGLTPEKKREYDLSKVGE


TFTMKAFMEADTLLSSIQRKSRKIIVHLTDGVPTRSYAINSFVKGSTYANQFERIKEKGYLDKNNYFITDDP


EKIKGNGESYFLFPLDSYQTQIISGNLQKLHYLDLNLNYPKGTIYRNGPVREHGTPTKLYINSLKQKNYDIF


NFGIDISGFRQVYNEDYKKNQDGTFQKLKEEAFELSDGEITELMNSFSSKPEYYTPIVTSADVSNNEILSKI


QQQFEKILTKENSIVNGTIEDPMGDKINLHLGNGQTLQPSDYTLQGNDGSIMKDSIATGGPNNDGGILKGVK


LEYIKNKLYVRGLNLGEGQKVTLTYDVKLDDSFISNKFYDTNGRTTLNPKSEEPDTLRDFPIPKIRDVREYP


TITIKNEKKLGEIEFTKVDKDNNKLLLKGATFELQEFNEDYKLYLPIKNNNSKVVTGENGKISYKDLKDGKY


QLIEAVSPKDYQKITNKPILTFEVVKGSIQNIIAVNKQISEYHEEGDKHLITNTHIPPKGIIPMTGGKGILS


FILIGGAMMSIAGGIYIWKRHKKSSDASIEKD


 


Strain BAA23


ORF DNA sequence (SEQ ID NO: 205):


ATGAGAAAATACCAAAAATTTTCTAAAATATTGACGTTAAGTCTTTTTTGTTTGTCGCAAATACCGCTTAAT


ACCAATGTTTTAGGGGAAAGTACCGTACCGGAAAATGGTGCTAAAGGAAAGTTAGTTGTTAAAAAGACAGAT


GACCAGAACAAACCACTTTCAAAAGCTACCTTTGTTTTAAAAACTACTGCTCATCCAGAAAGTAAAATAGAA


AAAGTAACTGCTGAGCTAACAGGTGAAGCTACTTTTGATAATCTCATACCTGGAGATTATACTTTATCAGAA


GAAACAGCGCCCGAAGGTTATAAAAAGACTAACCAGACTTGGCAAGTTAAGGTTGAGAGTAATGGAAAAACT


ACGATACAAAATAGTGGTGATAAAAATTCCACAATTGGACAAAATCAGGAAGAACTAGATAAGCAGTATCCC


CCCACAGGAATTTATGAAGATACAAAGGAATCTTATAAACTTGAGCATGTTAAAGGTTCAGTTCCAAATGGA


AAGTCAGAGGCAAAAGCAGTTAACCCATATTCAAGTGAAGGTGAGCATATAAGAGAAATTCCAGAGGGAACA


TTATCTAAACGTATTTCAGAAGTAGGTGATTTAGCTCATAATAAATATAAAATTGAGTTAACTGTCAGTGGA


AAAACCATAGTAAAACCAGTGGACAAACAAAAGCCGTTAGATGTTGTCTTCGTACTCGATAATTCTAACTCA


ATGAATAACGATGGCCCAAATTTTCAAAGGCATAATAAAGCCAAGAAAGCTGCCGAAGCTCTTGGGACCGCA


GTAAAAGATATTTTAGGAGCAAACAGTGATAATAGGGTTGCATTAGTTACCTATGGTTCAGATATTTTTGAT


GGTAGGAGTGTAGATGTCGTAAAAGGATTTAAAGAAGATGATAAATATTATGGCCTTCAAACTAAGTTCACA


ATTCAGACAGAGAATTATAGTCATAAACAATTAACAAATAATGCTGAAGAGATTATAAAAAGGATTCCGACA


GAAGCTCCTAAAGCTAAGTGGGGATCTACTACCAATGGATTAACTCCAGAGCAACAAAAGGAGTACTATCTT


AGTAAAGTAGGAGAAACATTTACTATGAAAGCCTTCATGGAGGCAGATGATATTTTGAGTCAAGTAAATCGA


AATAGTCAAAAAATTATTGTTCATGTAACTGATGGTGTTCCTACGAGATCATATGCTATTAATAATTTTAAA


CTGGGTGCATCATATGAAAGCCAATTTGAACAAATGAAAAAAAATGGATATCTAAATAAAAGTAATTTTCTA


CTTACTGATAAGCCCGAGGATATAAAAGGAAATGGGGAGAGTTACTTTTTGTTTCCCTTAGATAGTTATCAA


ACACAGATAATCTCTGGAAACTTACAAAAACTTCATTATTTAGATTTAAATCTTAATTACCCTAAAGGTACA


TTTTATCGAAATGGACCAGTAAGAGAACATGGAACACCAACCAAACTTTATATAAATAGTTTAAAACAGAAA


AATTATGACATCTTTAATTTTGGTATAGATATATCTGGTTTTAGACAAGTTTATAATGAGGATTATAAGAAA


AATCAAGATGGTACTTTTCAAAAATTGAAAGAGGAAGCTTTTGAACTTTCAGATGGGGAAATAACAGAACTA


ATGAAGTCATTCTCTTCTAAACCTGAGTATTATACCCCGATAGTAACTTCATCCGATGCATCTAACAATGAA


ATTTTATCTAAAATTCAGCAACAATTTGAAAAGATTTTAACAAAAGAAAACTCAATTGTTAATGGAACTATA


GAAGATCCTATGGGTGACAAAATCAATTTACAGCTTGGCAACGGACAAACATTGCAACCAAGTGATTATACT


TTACAGGGAAATGATGGAAGTATAATGAAAGATAGCATTGCAACTGGTGGGCCTAATAATGATGGTGGAATA


CTTAAAGGGGTTAAATTAGAATACATCAAAAATAAACTCTACGTTAGAGGTTTGAACTTAGGGGAGGGACAA


AAAGTAACACTCACATATGATGTGAAACTAGATGACAGTTTTATAAGTAACAAATTCTATGACACTAATGGT


AGAACAACATTGAATCCTAAATCAGAGGATCCTAATACACTTAGAGATTTTCCAATCCCTAAAATTCGTGAT


GTGAGAGAATATCCTACAATAACGATTAAAAACGAGAAGAAGTTAGGTGAAATTGAATTTACAAAAGTTGAT


AAAGATAATAATAAGTTGCTTCTCAAAGGAGCTACGTTTGAACTTCAAGAATTTAATGAAGATTATAAACTT


TATTTACCAATAAAAAATAATAATTCAAAAGTAGTGACGGGAGAAAACGGCAAAATTTCTTACAAAGATTTG


AAAGATGGCAAATATCAGTTAATAGAAGCAGTTTCGCCGAAGGATTATCAAAAAATTACTAATAAACCAATT


TTAACTTTTGAAGTTGTTAAAGGATCGATACAAAATATAATAGCTGTTAATAAACAGATTTCTGAATATCAT


GAGGAAGGTGACAAGCATTTAATTACCAACACGCATATTCCACCAAAAGGAATTATTCCGATGACAGGTGGG


AAAGGAATTCTATCTTTCATTTTAATAGGTGGATCTATGATGTCTATTGCAGGTGGAATTTATATTTGGAAA


AGATATAAGAAATCTAGTGATATATCTAGAGAAAAAGATTAA


 


ORF amino acid sequence (SEQ ID NO: 186):


MRKYQKFSKILTLSLFCLSQIPLNTNVLGESTVPENGAKGKLVVKKTDDQNKPLSKATFVLKTTAHPESKIE


KVTAELTGEATFDNLIPGDYTLSEETAPEGYKKTNQTWQVKVESNGKTTIQNSGDKNSTIGQNQEELDKQYP


PTGIYEDTKESYKLEHVKGSVPNGKSEAKAVNPYSSEGEHIREIPEGTLSKRISEVGDLAHNKYKIELTVSG


KTIVKPVDKQKPLDVVFVLDNSNSMNNDGPNFQRHNKAKKAAEALGTAVKDILGANSDNRVALVTYGSDIFD


GRSVDVVKGFKEDDKYYGLQTKFTIQTENYSHKQLTNNAEEIIKRIPTEAPKAKWGSTTNGLTPEQQKEYYL


SKVGETFTMKAFMEADDILSQVNRNSQKIIVHVTDGVPTRSYAINNFKLGASYESQFEQMKKNGYLNKSNFL


LTDKPEDIKGNGESYFLFPLDSYQTQIISGNLQKLHYLDLNLNYPKGTFYRNGPVREHGTPTKLYINSLKQK


NYDIFNFGIDISGFRQVYNEDYKKNQDGTFQKLKEEAFELSDGEITELMKSFSSKPEYYTPIVTSSDASNNE


ILSKIQQQFEKILTKENSIVNGTIEDPMGDKINLQLGNGQTLQPSDYTLQGNDGSIMKDSIATGGPNNDGGI


LKGVKLEYIKNKLYVRGLNLGEGQKVTLTYDVKLDDSFISNKFYDTNGRTTLNPKSEDPNTLRDFPIPKIRD


VREYPTITIKNEKKLGEIEFTKVDKDNNKLLLKGATFELQEFNEDYKLYLPIKNNNSKVVTGENGKISYKDL


KDGKYQLIEAVSPKDYQKITNKPILTFEVVKGSIQNIIAVNKQISEYHEEGDKHLITNTHIPPKGIIPMTGG


KGILSFILIGGSMMSIAGGIYIWKRYKKSSDISREKD


 


Strain IC98


ORF DNA sequence (SEQ ID NO: 206):


ATGAAAAAGAGACAAAAAATATGGAGAGGGTTATCAGTTACTTTACTAATCCTGTCCCAAATTCCATTTGGT


ATATTGGTACAAGGTGAAACCCAAGATACCAATCAAGCACTTGGAAAAGTAATTGTTAAAAAAACGGGAGAC


AATGCTACACCATTAGGCAAAGCGACTTTTGTGTTAAAAAATGACAATGATAAGTCAGAAACAAGTCACGAA


ACGGTAGAGGGTTCTGGAGAAGCAACCTTTGAAAACATAAAACCTGGAGACTACACATTAAGAGAAGAAACA


GCACCAATTGGTTATAAAAAAACTGATAAAACCTGGAAAGTTAAAGTTGCAGATAACGGAGCAACAATAATC


GAGGGTATGGATGCAGATAAAGCAGAGAAACGAAAAGAAGTTTTGAATGCCCAATATCCAAAATCAGCTATT


TATGAGGATACAAAAGAAAATTACCCATTAGTTAATGTAGAGGGTTCCAAAGTTGGTGAACAATACAAAGCA


TTGAATCCAATAAATGGAAAAGATGGTCGAAGAGAGATTGCTGAAGGTTGGTTATCAAAAAAAATTACAGGG


GTCAATGATCTCGATAAGAATAAATATAAAATTGAATTAACTGTTGAGGGTAAAACCACTGTTGAAACGAAA


GAACTTAATCAACCACTAGATGTCGTTGTGCTATTAGATAATTCAAATAGTATGAATAATGAAAGAGCCAAT


AATTCTCAAAGAGCATTAAAAGCTGGGGAAGCAGTTGAAAAGCTGATTGATAAAATTACATCAAATAAAGAC


AATAGAGTAGCTCTTGTGACATATGCCTCAACCATTTTTGATGGTACTGAAGCGACCGTATCAAAGGGAGTT


GCCGATCAAAATGGTAAAGCGCTGAATGATAGTGTATCATGGGATTATCATAAAACTACTTTTACAGCAACT


ACACATAATTACAGTTATTTAAATTTAACAAATGATGCTAACGAAGTTAATATTCTAAAGTCAAGAATTCCA


AAGGAAGCGGAGCATATAAATGGGGATCGCACGCTCTATCAATTTGGTGCGACATTTACTCAAAAAGCTCTA


ATGAAAGCAAATGAAATTTTAGAGACACAAAGTTCTAATGCTAGAAAAAAACTTATTTTTCACGTAACTGAT


GGTGTCCCTACGATGTCTTATGCCATAAATTTTAATCCTTATATATCAACATCTTACCAAAACCAGTTTAAT


TCTTTTTTAAATAAAATACCAGATAGAAGTGGTATTCTCCAAGAGGATTTTATAATCAATGGTGATGATTAT


CAAATAGTAAAAGGAGATGGAGAGAGTTTTAAACTGTTTTCGGATAGAAAAGTTCCTGTTACTGGAGGAACG


ACACAAGCAGCTTATCGAGTACCGCAAAATCAACTCTCTGTAATGAGTAATGAGGGATATGCAATTAATAGT


GGATATATTTATCTCTATTGGAGAGATTACAACTGGGTCTATCCATTTGATCCTAAGACAAAGAAAGTTTCT


GCAACGAAACAAATCAAAACTCATGGTGAGCCAACAACATTATACTTTAATGGAAATATAAGACCTAAAGGT


TATGACATTTTTACTGTTGGGATTGGTGTAAACGGAGATCCTGGTGCAACTCCTCTTGAAGCTGAGAAATTT


ATGCAATCAATATCAAGTAAAACAGAAAATTATACTAATGTTGATGATACAAATAAAATTTATGATGAGCTA


AATAAATACTTTAAAACAATTGTTGAGGAAAAACATTCTATTGTTGATGGAAATGTGACTGATCCTATGGGA


GAGATGATTGAATTCCAATTAAAAAATGGTCAAAGTTTTACACATGATGATTACGTTTTGGTTGGAAATGAT


GGCAGTCAATTAAAAAATGGTGTGGCTCTTGGTGGACCAAACAGTGATGGGGGAATTTTAAAAGATGTTACA


GTGACTTATGATAAGACATCTCAAACCATCAAAATCAATCATTTGAACTTAGGAAGTGGACAAAAAGTAGTT


CTTACCTATGATGTACGTTTAAAAGATAACTATATAAGTAACAAATTTTACAATACAAATAATCGTACAACG


CTAAGTCCGAAGAGTGAAAAAGAACCAAATACTATTCGTGATTTCCCAATTCCCAAAATTCGTGATGTTCGT


GAGTTTCCGGTACTAACCATCAGTAATCAGAAGAAAATGGGTGAGGTTGAATTTATTAAAGTTAATAAAGAC


AAACATTCAGAATCGCTTTTGGGAGCTAAGTTTCAACTTCAGATAGAAAAAGATTTTTCTGGGTATAAGCAA


TTTGTTCCAGAGGGAAGTGATGTTACAACAAAGAATGATGGTAAAATTTATTTTAAAGCACTTCAAGATGGT


AACTATAAATTATATGAAATTTCAAGTCCAGATGGCTATATAGAGGTTAAAACGAAACCTGTTGTGACATTT


ACAATTCAAAATGGAGAAGTTACGAACCTGAAAGCAGATCCAAATGCTAATAAAAATCAAATCGGGTATCTT


GAAGGAAATGGTAAACATCTTATTACCAACACTCCCAAACGCCCACCAGGTGTTTTTCCTAAAACAGGGGGA


ATTGGTACAATTGTCTATATATTAGTTGGTTCTACTTTTATGATACTTACCATTTGTTCTTTCCGTCGTAAA


CAATTGTAA


 


ORF amino acid sequence (SEQ ID NO: 187):


MKKRQKIWRGLSVTLLILSQIPFGILVQGETQDTNQALGKVIVKKTGDNATPLGKATFVLKNDNDKSETSHE


TVEGSGEATFENIKPGDYTLREETAPIGYKKTDKTWKVKVADNGATIIEGMDADKAEKRKEVLNAQYPKSAI


YEDTKENYPLVNVEGSKVGEQYKALNPINGKDGRREIAEGWLSKKITGVNDLDKNKYKIELTVEGKTTVETK


ELNQPLDVVVLLDNSNSMNNERANNSQRALKAGEAVEKLIDKITSNKDNRVALVTYASTIFDGTEATVSKGV


ADQNGKALNDSVSWDYHKTTFTATTHNYSYLNLTNDANEVNILKSRIPKEAEHINGDRTLYQFGATFTQKAL


MKANEILETQSSNARKKLIFHVTDGVPTMSYAINFNPYISTSYQNQFNSFLNKIPDRSGILQEDFIINGDDY


QIVKGDGESFKLFSDRKVPVTGGTTQAAYRVPQNQLSVMSNEGYAINSGYIYLYWRDYNWVYPFDPKTKKVS


ATKQIKTHGEPTTLYFNGNIRPKGYDIFTVGIGVNGDPGATPLEAEKFMQSISSKTENYTNVDDTNKIYDEL


NKYFKTIVEEKHSIVDGNVTDPMGEMIEFQLKNGQSFTHDDYVLVGNDGSQLKNGVALGGPNSDGGILKDVT


VTYDKTSQTIKINHLNLGSGQKVVLTYDVRLKDNYISNKFYNTNNRTTLSPKSEKEPNTIRDFPIPKIRDVR


EFPVLTISNQKKMGEVEFIKVNKDKHSESLLGAKFQLQIEKDFSGYKQFVPEGSDVTTKNDGKIYFKALQDG


NYKLYEISSPDGYIEVKTKPVVTFTIQNGEVTNLKADPNANKNQIGYLEGNGKHLITNTPKRPPGVFPKTGG


IGTIVYILVGSTFMILTICSFRRKQL


 


Strain IC105


ORF DNA sequence (SEQ ID NO: 207):


ATGAGAAAATACCAAAAATTTTCTAAAATATTGACGTTAAGTCTTTTTTGTTTGTCGCAAATACCGCTTAAT


ACCAATGTTTTAGGGGAAAGTACCGTACCGGAAAATGGTGCTAAAGGAAAGTTAGTTGTTAAAAAGACAGAT


GACCAGAACAAACCACTTTCAAAAGCTACCTTTGTTTTAAAAACTACTGCTCATCCAGAAAGTAAAATAGAA


AAAGTAACTGCTGAGGTAACAGGTGAAGCTACTTTTGATAATCTCACACCTGGAGATTACACTTTATCAGAA


GAAACGGCACCCGAAGGATACAAAAAGACTACCCAGACTTGGCAAGTTAAGGTTGAGAGTAATGGAAAAACT


ACGATACAAAATAGTGATGATAAAAAATCTATAATTGAACAAAGGCAAGAGGAACTAGATAAGCAGTATCCC


CTTACAGGAGCTTATGAAGATACAAAAGAATCTTATAATCTTGAGCATGTTAAAAATTCAATTCCAAATGGG


AAATTAGAGGCAAAAGCAGTTAATCCATATTCAAGTGAAGGTGAGCACATAAGAGAAATTCAAGAGGGAACA


TTATCTAAACGTATTTCAGAAGTAAATGATTTGGATCATAATAAATATAAAATTGAGTTAACTGTTAGCGGT


AAATCCATAATAAAAACTATAAATAAAGATGAACCTCTGGATGTTGTTTTTGTTCTTGATAATTCAAATTCT


ATGAAGAATAATGGAAAAAATAACAAGGCAAAAAAGGCAGGTGAAGCAGTAGAAACAATTATAAAAGATGTT


TTAGGAGCAAATGTTGAAAACCGAGCAGCTTTAGTTACTTATGGTTCAGATATTTTTGATGGAAGGACAGTT


AAAGTTATAAAAGGTTTTAAAGAGGATCCTTATTATGGACTTGAAACTAGTTTCACAGTTCAGACAAATGAT


TATAGCTATAAAAAGTTCACTAATATTGCTGCTGATATTATAAAAAAGATCCCTAAAGAAGCTCCAGAAGCT


AAGTGGGGGGGGACAAGTCTAGGATTAACTCCAGAAAAAAAGAGGGAATATGATTTAAGTAAAGTAGGTGAG


ACCTTTACAATGAAAGCTTTTATGGAGGCAGATACCTTGTTAAGTAGTATACAGCGTAAGAGTAGAAAGATT


ATTGTTCATCTAACTGACGGTGTTCCAACAAGATCATATGCCATTAATAGTTTTGTAACAGGTTCAACATAC


GCAAATCAATTTGAGAGAATAAAAGAAAAAGGTTATTTAGACAAAAATAATTATTTTATAACTGATGATCCA


GAAAAGATCAAAGGCAATGGGGAGAGTTACTTTTTGTTTCCCTTAGATAGTTATCAAACACAGATAATTTCT


GGAAACTTACAAAAACTTCATTATTTAGATTTAAATCTTAATTACCCTAAAGGTACAATTTATAGAAATGGA


CCAGTAAGAGAACATGGAACACCAACCAAACTTTATATAAATAGTTTAAAACAGAAAAATTATGACATCTTT


AATTTTGGTATAGATATATCTGGTTTTAGACAAGTTTATAATGAGGATTATAAGAAAAATCAAGATGGTACT


TTTCAAAAATTGAAGGAGGAAGCTTTTGAACTTTCAGATGGGGAAATAACAGAACTAATGAATTCATTCTCT


TCTAAACCTGAGTATTATACCCCGATAGTAACTTCAGCTGATGTATCTAATAATGAAATTTTATCTAAAATT


CAGCAACAATTTGAAAAGATTTTAACAAAGGAAAACTCAATTGTTAATGGAACTATAGAAGATCCTATGGGT


GATAAAATCAATTTACATCTTGGCAACGGACAAACATTGCAACCAAGTGATTATACTTTACAGGGAAATGAT


GGAAGTATAATGAAAGATAGCATTGCAACTGGAGGGCCTAATAATGATGGTGGGATACTTAAAGGGGTTAAA


TTAGAATACATCAAAAATAAACTCTACGTTAGAGGTTTGAACTTAGGGGAGGGGCAAAAAGTAACACTCACA


TATGATGTGAAACTAGATGACAGTTTTATTAGTAACAAATTCTATGACACTAATGGTAGAACAACATTGAAT


CCTAAATCAGAGGAACCCGATACACTTAGAGATTTTCCAATCCCTAAAATTCGTGATGTGAGAGAATATCCT


ACAATAACGATTAAAAACGAGAAGAAGTTAGGTGAAATTGAATTTACAAAAGTTGATAAAGATAATAATAAG


TTGCTTCTCAAAGGAGCTACATTTGAACTTCAAGAATTTAATGAAGATTATAAACTTTATTTACCAATAAAA


AATAATAATTCAAAAGTAGTGACGGGAGAAAACGGCAAAATTTCTTACAAAGATTTGAAAGATGGCAAATAT


CAGTTAATAGAAGCAGTTTCGCCGAAGGATTATCAAAAAATTACTAATAAACCAATTTTAACTTTTGAAGTT


GTTAAAGGATCGATACAAAATATAATAGCTGTTAATAAACAGATTTCTGAATATCATGAGGAAGGTGACAAG


CATTTAATTACCAACACGCATATTCCACCAAAAGGAATTATTCCGATGACAGGTGGGAAAGGAATTCTATCT


TTCATTTTAATAGGTGGAGCTATGATGTCTATTGCAGGTGGAATTTATATTTGGAAAAGACATAAGAAATCT


AGTGATGCATCAATCGAGAAAGATTAA


 


ORF amino acid sequence (SEQ ID NO: 188):


MRKYQKFSKILTLSLFCLSQIPLNTNVLGESTVPENGAKGKLVVKKTDDQNKPLSKATFVLKTTAHPESKIE


KVTAEVTGEATFDNLTPGDYTLSEETAPEGYKKTTQTWQVKVESNGKTTIQNSDDKKSIIEQRQEELDKQYP


LTGAYEDTKESYNLEHVKNSIPNGKLEAKAVNPYSSEGEHIREIQEGTLSKRISEVNDLDHNKYKIELTVSG


KSIIKTINKDEPLDVVFVLDNSNSMKNNGKNNKAKKAGEAVETIIKDVLGANVENRAALVTYGSDIFDGRTV


KVIKGFKEDPYYGLETSFTVQTNDYSYKKFTNIAADIIKKIPKEAPEAKWGGTSLGLTPEKKREYDLSKVGE


TFTMKAFMEADTLLSSIQRKSRKIIVHLTDGVPTRSYAINSFVTGSTYANQFERIKEKGYLDKNNYFITDDP


EKIKGNGESYFLFPLDSYQTQIISGNLQKLHYLDLNLNYPKGTIYRNGPVREHGTPTKLYINSLKQKNYDIF


NFGIDISGFRQVYNEDYKKNQDGTFQKLKEEAFELSDGEITELMNSFSSKPEYYTPIVTSADVSNNEILSKI


QQQFEKILTKENSIVNGTIEDPMGDKINLHLGNGQTLQPSDYTLQGNDGSIMKDSIATGGPNNDGGILKGVK


LEYIKNKLYVRGLNLGEGQKVTLTYDVKLDDSFISNKFYDTNGRTTLNPKSEEPDTLRDFPIPKIRDVREYP


TITIKNEKKLGEIEFTKVDKDNNKLLLKGATFELQEFNEDYKLYLPIKNNNSKVVTGENGKISYKDLKDGKY


QLIEAVSPKDYQKITNKPILTFEVVKGSIQNIIAVNKQISEYHEEGDKHLITNTHIPPKGIIPMTGGKGILS


FILIGGAMMSIAGGIYIWKRHKKSSDASIEKD


 


Strain IC108


ORF DNA sequence (SEQ ID NO: 208):


ATGAAAAAGAGACAAAAAATATGGAGAGGGTTATCAGTTACTTTACTAATCCTGTCCCAAATTCCATTTGGT


ATATTGGTACAAGGTGAAACCCAAGATACCAATCAAGCACTTGGAAAAGTAATTGTTAAAAAAACGGGAGAC


AATGCTACACCATTAGGCAAAGCGACTTTTGTGTTAAAAAATGACAATGATAAGTCAGAAACAAGTCACGAA


ACGGTAGAGGGTTCTGGAGAAGCAACCTTTGAAAACATAAAACCTGGAGACTACACATTAAGAGAAGAAACA


GCACCAATTGGTTATAAAAAAACTGATAAAACCTGGAAAGTTAAAGTTGCAGATAACGGAGCAACAATAATC


GAGGGTATGGATGCAGATAAAGCAGAGAAACGAAAAGAAGTTTTGAATGCCCAATATCCAAAATCAGCTATT


TATGAGGATACAAAAGAAAATTACCCATTAGTTAATGTAGAGGGTTCCAAAGTTGGTGAACAATACAAAGCA


TTGAATCCAATAAATGGAAAAGATGGTCGAAGAGAGATTGCTGAAGGTTGGTTATCAAAAAAAATTACAGGG


GTCAATGATCTCGATAAGAATAAATATAAAATTGAATTAACTGTTGAGGGTAAAACCACTGTTGAAACGAAA


GAACTTAATCAACCACTAGATGTCGTTGTGCTATTAGATAATTCAAATAGTATGAATAATGAAAGAGCCAAT


AATTCTCAAAGAGCATTAAAAGCTGGGGAAGCAGTTGAAAAGCTGATTGATAAAATTACATCAAATAAAGAC


AATAGAGTAGCTCTTGTGACATATGCCTCAACCATTTTTGATGGTACTGAAGCGACCGTATCAAAGGGAGTT


GCCGATCAAAATGGTAAAGCGCTGAATGATAGTGTATCATGGGATTATCATAAAACTACTTTTACAGCAACT


ACACATAATTACAGTTATTTAAATTTAACAAATGATGCTAACGAAGTTAATATTCTAAAGTCAAGAATTCCA


AAGGAAGCGGAGCATATAAATGGGGATCGCACGCTCTATCAATTTGGTGCGACATTTACTCAAAAAGCTCTA


ATGAAAGCAAATGAAATTTTAGAGACACAAAGTTCTAATGCTAGAAAAAAACTTATTTTTCACGTAACTGAT


GGTGTCCCTACGATGTCTTATGCCATAAATTTTAATCCTTATATATCAACATCTTACCAAAACCAGTTTAAT


TCTTTTTTAAATAAAATACCAGATAGAAGTGGTATTCTCCAAGAGGATTTTATAATCAATGGTGATGATTAT


CAAATAGTAAAAGGAGATGGAGAGAGTTTTAAACTGTTTTCGGATAGAAAAGTTCCTGTTACTGGAGGAACG


ACACAAGCAGCTTATCGAGTACCGCAAAATCAACTCTCTGTAATGAGTAATGAGGGATATGCAATTAATAGT


GGATATATTTATCTCTATTGGAGAGATTACAACTGGGTCTATCCATTTGATCCTAAGACAAAGAAAGTTTCT


GCAACGAAACAAATCAAAACTCATGGTGAGCCAACAACATTATACTTTAATGGAAATATAAGACCTAAAGGT


TATGACATTTTTACTGTTGGGATTGGTGTAAACGGAGATCCTGGTGCAACTCCTCTTGAAGCTGAGAAATTT


ATGCAATCAATATCAAGTAAAACAGAAAATTATACTAATGTTGATGATACAAATAAAATTTATGATGAGCTA


AATAAATACTTTAAAACAATTGTTGAGGAAAAACATTCTATTGTTGATGGAAATGTGACTGATCCTATGGGA


GAGATGATTGAATTCCAATTAAAAAATGGTCAAAGTTTTACACATGATGATTACGTTTTGGTTGGAAATGAT


GGCAGTCAATTAAAAAATGGTGTGGCTCTTGGTGGACCAAACAGTGATGGGGGAATTTTAAAAGATGTTACA


GTGACTTATGATAAGACATCTCAAACCATCAAAATCAATCATTTGAACTTAGGAAGTGGACAAAAAGTAGTT


CTTACCTATGATGTACGTTTAAAAGATAACTATATAAGTAACAAATTTTACAATACAAATAATCGTACAACG


CTAAGTCCGAAGAGTGAAAAAGAACCAAATACTATTCGTGATTTCCCAATTCCCAAAATTCGTGATGTTCGT


GAGTTTCCGGTACTAACCATCAGTAATCAGAAGAAAATGGGTGAGGTTGAATTTATTAAAGTTAATAAAGAC


AAACATTCAGAATCGCTTTTGGGAGCTAAGTTTCAACTTCAGATAGAAAAAGATTTTTCTGGGTATAAGCAA


TTTGTTCCAGAGGGAAGTGATGTTACAACAAAGAATGATGGTAAAATTTATTTTAAAGCACTTCAAGATGGT


AACTATAAATTATATGAAATTTCAAGTCCAGATGGCTATATAGAGGTTAAAACGAAACCTGTTGTGACATTT


ACAATTCAAAATGGAGAAGTTACGAACCTGAAAGCAGATCCAAATGCTAATAAAAATCAAATCGGGTATCTT


GAAGGAAATGGTAAACATCTTATTACCAACACTCCCAAACGCCCACCAGGTGTTTTTCCTAAAACAGGGGGA


ATTGGTACAATTGTCTATATATTAGTTGGTTCTACTTTTATGATACTTACCATTTGTTCTTTCCGTCGTAAA


CAATTGTAA


 


ORF amino acid sequence (SEQ ID NO: 189):


MKKRQKIWRGLSVTLLILSQIPFGILVQGETQDTNQALGKVIVKKTGDNATPLGKATFVLKNDNDKSETSHE


TVEGSGEATFENIKPGDYTLREETAPIGYKKTDKTWKVKVADNGATIIEGMDADKAEKRKEVLNAQYPKSAI


YEDTKENYPLVNVEGSKVGEQYKALNPINGKDGRREIAEGWLSKKITGVNDLDKNKYKIELTVEGKTTVETK


ELNQPLDVVVLLDNSNSMNNERANNSQRALKAGEAVEKLIDKITSNKDNRVALVTYASTIFDGTEATVSKGV


ADQNGKALNDSVSWDYHKTTFTATTHNYSYLNLTNDANEVNILKSRIPKEAEHINGDRTLYQFGATFTQKAL


MKANEILETQSSNARKKLIFHVTDGVPTMSYAINFNPYISTSYQNQFNSFLNKIPDRSGILQEDFIINGDDY


QIVKGDGESFKLFSDRKVPVTGGTTQAAYRVPQNQLSVMSNEGYAINSGYIYLYWRDYNWVYPFDPKTKKVS


ATKQIKTHGEPTTLYFNGNIRPKGYDIFTVGIGVNGDPGATPLEAEKFMQSISSKTENYTNVDDTNKIYDEL


NKYFKTIVEEKHSIVDGNVTDPMGEMIEFQLKNGQSFTHDDYVLVGNDGSQLKNGVALGGPNSDGGILKDVT


VTYDKTSQTIKINHLNLGSGQKVVLTYDVRLKDNYISNKFYNTNNRTTLSPKSEKEPNTIRDFPIPKIRDVR


EFPVLTISNQKKMGEVEFIKVNKDKHSESLLGAKFQLQIEKDFSGYKQFVPEGSDVTTKNDGKIYFKALQDG


NYKLYEISSPDGYIEVKTKPVVTFTIQNGEVTNLKADPNANKNQIGYLEGNGKHLITNTPKRPPGVFPKTGG


IGTIVYILVGSTFMILTICSFRRKQL


 


Strain IC216


ORF DNA sequence (SEQ ID NO: 209):


ATGAAAAAGAGACAAAAAATATGGAGAGGGTTATCAGTTACTTTACTAATCCTGTCCCAAATTCCATTTGGT


ATATTGGTACAAGGTGAAACCCAAGATACCAATCAAGCACTTGGAAAAGTAATTGTTAAAAAAACGGGAGAC


AATGCTACACCATTAGGCAAAGCGACTTTTGTGTTAAAAAATGACAATGATAAGTCAGAAACAAGTCACGAA


ACGGTAGAGGGTTCTGGAGAAGCAACCTTTGAAAACATAAAACCTGGAGACTACACATTAAGAGAAGAAACA


GCACCAATTGGTTATAAAAAAACTGATAAAACCTGGAAAGTTAAAGTTGCAGATAACGGAGCAACAATAATC


GAGGGTATGGATGCAGATAAAGCAGAGAAACGAAAAGAAGTTTTGAATGCCCAATATCCAAAATCAGCTATT


TATGAGGATACAAAAGAAAATTACCCATTAGTTAATGTAGAGGGTTCCAAAGTTGGTGAACAATACAAAGCA


TTGAATCCAATAAATGGAAAAGATGGTCGAAGAGAGATTGCTGAAGGTTGGTTATCAAAAAAAATTACAGGG


GTCAATGATCTCGATAAGAATAAATATAAAATTGAATTAACTGTTGAGGGTAAAACCACTGTTGAAACGAAA


GAACTTAATCAACCACTAGATGTCGTTGTGCTATTAGATAATTCAAATAGTATGAATAATGAAAGAGCCAAT


AATTCTCAAAGAGCATTAAAAGCTGGGGAAGCAGTTGAAAAGCTGATTGATAAAATTACATCAAATAAAGAC


AATAGAGTAGCTCTTGTGACATATGCCTCAACCATTTTTGATGGTACTGAAGCGACCGTATCAAAGGGAGTT


GCCGATCAAAATGGTAAAGCGCTGAATGATAGTGTATCATGGGATTATCATAAAACTACTTTTACAGCAACT


ACACATAATTACAGTTATTTAAATTTAACAAATGATGCTAACGAAGTTAATATTCTAAAGTCAAGAATTCCA


AAGGAAGCGGAGCATATAAATGGGGATCGCACGCTCTATCAATTTGGTGCGACATTTACTCAAAAAGCTCTA


ATGAAAGCAAATGAAATTTTAGAGACACAAAGTTCTAATGCTAGAAAAAAACTTATTTTTCACGTAACTGAT


GGTGTCCCTACGATGTCTTATGCCATAAATTTTAATCCTTATATATCAACATCTTACCAAAACCAGTTTAAT


TCTTTTTTAAATAAAATACCAGATAGAAGTGGTATTCTCCAAGAGGATTTTATAATCAATGGTGATGATTAT


CAAATAGTAAAAGGAGATGGAGAGAGTTTTAAACTGTTTTCGGATAGAAAAGTTCCTGTTACTGGAGGAACG


ACACAAGCAGCTTATCGAGTACCGCAAAATCAACTCTCTGTAATGAGTAATGAGGGATATGCAATTAATAGT


GGATATATTTATCTCTATTGGAGAGATTACAACTGGGTCTATCCATTTGATCCTAAGACAAAGAAAGTTTCT


GCAACGAAACAAATCAAAACTCATGGTGAGCCAACAACATTATACTTTAATGGAAATATAAGACCTAAAGGT


TATGACATTTTTACTGTTGGGATTGGTGTAAACGGAGATCCTGGTGCAACTCCTCTTGAAGCTGAGAAATTT


ATGCAATCAATATCAAGTAAAACAGAAAATTATACTAATGTTGATGATACAAATAAAATTTATGATGAGCTA


AATAAATACTTTAAAACAATTGTTGAGGAAAAACATTCTATTGTTGATGGAAATGTGACTGATCCTATGGGA


GAGATGATTGAATTCCAATTAAAAAATGGTCAAAGTTTTACACATGATGATTACGTTTTGGTTGGAAATGAT


GGCAGTCAATTAAAAAATGGTGTGGCTCTTGGTGGACCAAACAGTGATGGGGGAATTTTAAAAGATGTTACA


GTGACTTATGATAAGACATCTCAAACCATCAAAATCAATCATTTGAACTTAGGAAGTGGACAAAAAGTAGTT


CTTACCTATGATGTACGTTTAAAAGATAACTATATAAGTAACAAATTTTACAATACAAATAATCGTACAACG


CTAAGTCCGAAGAGTGAAAAAGAACCAAATACTATTCGTGATTTCCCAATTCCCAAAATTCGTGATGTTCGT


GAGTTTCCGGTACTAACCATCAGTAATCAGAAGAAAATGGGTGAGGTTGAATTTATTAAAGTTAATAAAGAC


AAACATTCAGAATCGCTTTTGGGAGCTAAGTTTCAACTTCAGATAGAAAAAGATTTTTCTGGGTATAAGCAA


TTTGTTCCAGAGGGAAGTGATGTTACAACAAAGAATGATGGTAAAATTTATTTTAAAGCACTTCAAGATGGT


AACTATAAATTATATGAAATTTCAAGTCCAGATGGCTATATAGAGGTTAAAACGAAACCTGTTGTGACATTT


ACAATTCAAAATGGAGAAGTTACGAACCTGAAAGCAGATCCAAATGCTAATAAAAATCAAATCGGGTATCTT


GAAGGAAATGGTAAACATCTTATTACCAACACTCCCAAACGCCCACCAGGTGTTTTTCCTAAAACAGGGGGA


ATTGGTACAATTGTCTATATATTAGTTGGTTCTACTTTTATGATACTTACCATTTGTTCTTTCCGTCGTAAA


CAATTGTAA


 


ORF amino acid sequence (SEQ ID NO: 190):


MKKRQKIWRGLSVTLLILSQIPFGILVQGETQDTNQALGKVIVKKTGDNATPLGKATFVLKNDNDKSETSHE


TVEGSGEATFENIKPGDYTLREETAPIGYKKTDKTWKVKVADNGATIIEGMDADKAEKRKEVLNAQYPKSAI


YEDTKENYPLVNVEGSKVGEQYKALNPINGKDGRREIAEGWLSKKITGVNDLDKNKYKIELTVEGKTTVETK


ELNQPLDVVVLLDNSNSMNNERANNSQRALKAGEAVEKLIDKITSNKDNRVALVTYASTIFDGTEATVSKGV


ADQNGKALNDSVSWDYHKTTFTATTHNYSYLNLTNDANEVNILKSRIPKEAEHINGDRTLYQFGATFTQKAL


MKANEILETQSSNARKKLIFHVTDGVPTMSYAINFNPYISTSYQNQFNSFLNKIPDRSGILQEDFIINGDDY


QIVKGDGESFKLFSDRKVPVTGGTTQAAYRVPQNQLSVMSNEGYAINSGYIYLYWRDYNWVYPFDPKTKKVS


ATKQIKTHGEPTTLYFNGNIRPKGYDIFTVGIGVNGDPGATPLEAEKFMQSISSKTENYTNVDDTNKIYDEL


NKYFKTIVEEKHSIVDGNVTDPMGEMIEFQLKNGQSFTHDDYVLVGNDGSQLKNGVALGGPNSDGGILKDVT


VTYDKTSQTIKINHLNLGSGQKVVLTYDVRLKDNYISNKFYNTNNRTTLSPKSEKEPNTIRDFPIPKIRDVR


EFPVLTISNQKKMGEVEFIKVNKDKHSESLLGAKFQLQIEKDFSGYKQFVPEGSDVTTKNDGKIYFKALQDG


NYKLYEISSPDGYIEVKTKPVVTFTIQNGEVTNLKADPNANKNQIGYLEGNGKHLITNTPKRPPGVFPKTGG


IGTIVYILVGSTFMILTICSFRRKQL


 


Strain IC244


ORF DNA sequence (SEQ ID NO: 210):


ATGAAAAAGAGACAAAAAATATGGAGAGGGTTATCAGTTACTTTACTAATCCTGTCCCAAATTCCATTTGGT


ATATTGGTACAAGGTGAAACCCAAGATACCAATCAAGCACTTGGAAAAGTAATTGTTAAAAAAACGGGAGAC


AATGCTACACCATTAGGCAAAGCGACTTTTGTGTTAAAAAATGACAATGATAAGTCAGAAACAAGTCACGAA


ACGGTAGAGGGTTCTGGAGAAGCAACCTTTGAAAACATAAAACCTGGAGACTACACATTAAGAGAAGAAACA


GCACCAATTGGTTATAAAAAAACTGATAAAACCTGGAAAGTTAAAGTTGCAGATAACGGAGCAACAATAATC


GAGGGTATGGATGCAGATAAAGCAGAGAAACGAAAAGAAGTTTTGAATGCCCAATATCCAAAATCAGCTATT


TATGAGGATACAAAAGAAAATTACCCATTAGTTAATGTAGAGGGTTCCAAAGTTGGTGAACAATACAAAGCA


TTGAATCCAATAAATGGAAAAGATGGTCGAAGAGAGATTGCTGAAGGTTGGTTATCAAAAAAAATTACAGGG


GTCAATGATCTCGATAAGAATAAATATAAAATTGAATTAACTGTTGAGGGTAAAACCACTGTTGAAACGAAA


GAACTTAATCAACCACTAGATGTCGTTGTGCTATTAGATAATTCAAATAGTATGAATAATGAAAGAGCCAAT


AATTCTCAAAGAGCATTAAAAGCTGGGGAAGCAGTTGAAAAGCTGATTGATAAAATTACATCAAATAAAGAC


AATAGAGTAGCTCTTGTGACATATGCCTCAACCATTTTTGATGGTACTGAAGCGACCGTATCAAAGGGAGTT


GCCGATCAAAATGGTAAAGCGCTGAATGATAGTGTATCATGGGATTATCATAAAACTACTTTTACAGCAACT


ACACATAATTACAGTTATTTAAATTTAACAAATGATGCTAACGAAGTTAATATTCTAAAGTCAAGAATTCCA


AAGGAAGCGGAGCATATAAATGGGGATCGCACGCTCTATCAATTTGGTGCGACATTTACTCAAAAAGCTCTA


ATGAAAGCAAATGAAATTTTAGAGACACAAAGTTCTAATGCTAGAAAAAAACTTATTTTTCACGTAACTGAT


GGTGTCCCTACGATGTCTTATGCCATAAATTTTAATCCTTATATATCAACATCTTACCAAAACCAGTTTAAT


TCTTTTTTAAATAAAATACCAGATAGAAGTGGTATTCTCCAAGAGGATTTTATAATCAATGGTGATGATTAT


CAAATAGTAAAAGGAGATGGAGAGAGTTTTAAACTGTTTTCGGATAGAAAAGTTCCTGTTACTGGAGGAACG


ACACAAGCAGCTTATCGAGTACCGCAAAATCAACTCTCTGTAATGAGTAATGAGGGATATGCAATTAATAGT


GGATATATTTATCTCTATTGGAGAGATTACAACTGGGTCTATCCATTTGATCCTAAGACAAAGAAAGTTTCT


GCAACGAAACAAATCAAAACTCATGGTGAGCCAACAACATTATACTTTAATGGAAATATAAGACCTAAAGGT


TATGACATTTTTACTGTTGGGATTGGTGTAAACGGAGATCCTGGTGCAACTCCTCTTGAAGCTGAGAAATTT


ATGCAATCAATATCAAGTAAAACAGAAAATTATACTAATGTTGATGATACAAATAAAATTTATGATGAGCTA


AATAAATACTTTAAAACAATTGTTGAGGAAAAACATTCTATTGTTGATGGAAATGTGACTGATCCTATGGGA


GAGATGATTGAATTCCAATTAAAAAATGGTCAAAGTTTTACACATGATGATTACGTTTTGGTTGGAAATGAT


GGCAGTCAATTAAAAAATGGTGTGGCTCTTGGTGGACCAAACAGTGATGGGGGAATTTTAAAAGATGTTACA


GTGACTTATGATAAGACATCTCAAACCATCAAAATCAATCATTTGAACTTAGGAAGTGGACAAAAAGTAGTT


CTTACCTATGATGTACGTTTAAAAGATAACTATATAAGTAACAAATTTTACAATACAAATAATCGTACAACG


CTAAGTCCGAAGAGTGAAAAAGAACCAAATACTATTCGTGATTTCCCAATTCCCAAAATTCGTGATGTTCGT


GAGTTTCCGGTACTAACCATCAGTAATCAGAAGAAAATGGGTGAGGTTGAATTTATTAAAGTTAATAAAGAC


AAACATTCAGAATCGCTTTTGGGAGCTAAGTTTCAACTTCAGATAGAAAAAGATTTTTCTGGGTATAAGCAA


TTTGTTCCAGAGGGAAGTGATGTTACAACAAAGAATGATGGTAAAATTTATTTTAAAGCACTTCAAGATGGT


AACTATAAATTATATGAAATTTCAAGTCCAGATGGCTATATAGAGGTTAAAACGAAACCTGTTGTGACATTT


ACAATTCAAAATGGAGAAGTTACGAACCTGAAAGCAGATCCAAATGCTAATAAAAATCAAATCGGGTATCTT


GAAGGAAATGGTAAACATCTTATTACCAACACTCCCAAACGCCCACCAGGTGTTTTTCCTAAAACAGGGGGA


ATTGGTACAATTGTCTATATATTAGTTGGTTCTACTTTTATGATACTTACCATTTGTTCTTTCCGTCGTAAA


CAATTGTAA


 


ORF amino acid sequence (SEQ ID NO: 191):


MKKRQKIWRGLSVTLLILSQIPFGILVQGETQDTNQALGKVIVKKTGDNATPLGKATFVLKNDNDKSETSHE


TVEGSGEATFENIKPGDYTLREETAPIGYKKTDKTWKVKVADNGATIIEGMDADKAEKRKEVLNAQYPKSAI


YEDTKENYPLVNVEGSKVGEQYKALNPINGKDGRREIAEGWLSKKITGVNDLDKNKYKIELTVEGKTTVETK


ELNQPLDVVVLLDNSNSMNNERANNSQRALKAGEAVEKLIDKITSNKDNRVALVTYASTIFDGTEATVSKGV


ADQNGKALNDSVSWDYHKTTFTATTHNYSYLNLTNDANEVNILKSRIPKEAEHINGDRTLYQFGATFTQKAL


MKANEILETQSSNARKKLIFHVTDGVPTMSYAINFNPYISTSYQNQFNSFLNKIPDRSGILQEDFIINGDDY


QIVKGDGESFKLFSDRKVPVTGGTTQAAYRVPQNQLSVMSNEGYAINSGYIYLYWRDYNWVYPFDPKTKKVS


ATKQIKTHGEPTTLYFNGNIRPKGYDIFTVGIGVNGDPGATPLEAEKFMQSISSKTENYTNVDDTNKIYDEL


NKYFKTIVEEKHSIVDGNVTDPMGEMIEFQLKNGQSFTHDDYVLVGNDGSQLKNGVALGGPNSDGGILKDVT


VTYDKTSQTIKINHLNLGSGQKVVLTYDVRLKDNYISNKFYNTNNRTTLSPKSEKEPNTIRDFPIPKIRDVR


EFPVLTISNQKKMGEVEFIKVNKDKHSESLLGAKFQLQIEKDFSGYKQFVPEGSDVTTKNDGKIYFKALQDG


NYKLYEISSPDGYIEVKTKPVVTFTIQNGEVTNLKADPNANKNQIGYLEGNGKHLITNTPKRPPGVFPKTGG


IGTIVYILVGSTFMILTICSFRRKQL


 


Strain IC245


ORF DNA sequence (SEQ ID NO: 211):


ATGAAAAAGAGACAAAAAATATGGAGAGGGTTATCAGTTACTTTACTAATCCTGTCCCAAATTCCATTTGGT


ATATTGGTACAAGGTGAAACCCAAGATACCAATCAAGCACTTGGAAAAGTAATTGTTAAAAAAACGGGAGAC


AATGCTACACCATTAGGCAAAGCGACTTTTGTGTTAAAAAATGACAATGATAAGTCAGAAACAAGTCACGAA


ACGGTAGAGGGTTCTGGAGAAGCAACCTTTGAAAACATAAAACCTGGAGACTACACATTAAGAGAAGAAACA


GCACCAATTGGTTATAAAAAAACTGATAAAACCTGGAAAGTTAAAGTTGCAGATAACGGAGCAACAATAATC


GAGGGTATGGATGCAGATAAAGCAGAGAAACGAAAAGAAGTTTTGAATGCCCAATATCCAAAATCAGCTATT


TATGAGGATACAAAAGAAAATTACCCATTAGTTAATGTAGAGGGTTCCAAAGTTGGTGAACAATACAAAGCA


TTGAATCCAATAAATGGAAAAGATGGTCGAAGAGAGATTGCTGAAGGTTGGTTATCAAAAAAAATTACAGGG


GTCAATGATCTCGATAAGAATAAATATAAAATTGAATTAACTGTTGAGGGTAAAACCACTGTTGAAACGAAA


GAACTTAATCAACCACTAGATGTCGTTGTGCTATTAGATAATTCAAATAGTATGAATAATGAAAGAGCCAAT


AATTCTCAAAGAGCATTAAAAGCTGGGGAAGCAGTTGAAAAGCTGATTGATAAAATTACATCAAATAAAGAC


AATAGAGTAGCTCTTGTGACATATGCCTCAACCATTTTTGATGGTACTGAAGCGACCGTATCAAAGGGAGTT


GCCGATCAAAATGGTAAAGCGCTGAATGATAGTGTATCATGGGATTATCATAAAACTACTTTTACAGCAACT


ACACATAATTACAGTTATTTAAATTTAACAAATGATGCTAACGAAGTTAATATTCTAAAGTCAAGAATTCCA


AAGGAAGCGGAGCATATAAATGGGGATCGCACGCTCTATCAATTTGGTGCGACATTTACTCAAAAAGCTCTA


ATGAAAGCAAATGAAATTTTAGAGACACAAAGTTCTAATGCTAGAAAAAAACTTATTTTTCACGTAACTGAT


GGTGTCCCTACGATGTCTTATGCCATAAATTTTAATCCTTATATATCAACATCTTACCAAAACCAGTTTAAT


TCTTTTTTAAATAAAATACCAGATAGAAGTGGTATTCTCCAAGAGGATTTTATAATCAATGGTGATGATTAT


CAAATAGTAAAAGGAGATGGAGAGAGTTTTAAACTGTTTTCGGATAGAAAAGTTCCTGTTACTGGAGGAACG


ACACAAGCAGCTTATCGAGTACCGCAAAATCAACTCTCTGTAATGAGTAATGAGGGATATGCAATTAATAGT


GGATATATTTATCTCTATTGGAGAGATTACAACTGGGTCTATCCATTTGATCCTAAGACAAAGAAAGTTTCT


GCAACGAAACAAATCAAAACTCATGGTGAGCCAACAACATTATACTTTAATGGAAATATAAGACCTAAAGGT


TATGACATTTTTACTGTTGGGATTGGTGTAAACGGAGATCCTGGTGCAACTCCTCTTGAAGCTGAGAAATTT


ATGCAATCAATATCAAGTAAAACAGAAAATTATACTAATGTTGATGATACAAATAAAATTTATGATGAGCTA


AATAAATACTTTAAAACAATTGTTGAGGAAAAACATTCTATTGTTGATGGAAATGTGACTGATCCTATGGGA


GAGATGATTGAATTCCAATTAAAAAATGGTCAAAGTTTTACACATGATGATTACGTTTTGGTTGGAAATGAT


GGCAGTCAATTAAAAAATGGTGTGGCTCTTGGTGGACCAAACAGTGATGGGGGAATTTTAAAAGATGTTACA


GTGACTTATGATAAGACATCTCAAACCATCAAAATCAATCATTTGAACTTAGGAAGTGGACAAAAAGTAGTT


CTTACCTATGATGTACGTTTAAAAGATAACTATATAAGTAACAAATTTTACAATACAAATAATCGTACAACG


CTAAGTCCGAAGAGTGAAAAAGAACCAAATACTATTCGTGATTTCCCAATTCCCAAAATTCGTGATGTTCGT


GAGTTTCCGGTACTAACCATCAGTAATCAGAAGAAAATGGGTGAGGTTGAATTTATTAAAGTTAATAAAGAC


AAACATTCAGAATCGCTTTTGGGAGCTAAGTTTCAACTTCAGATAGAAAAAGATTTTTCTGGGTATAAGCAA


TTTGTTCCAGAGGGAAGTGATGTTACAACAAAGAATGATGGTAAAATTTATTTTAAAGCACTTCAAGATGGT


AACTATAAATTATATGAAATTTCAAGTCCAGATGGCTATATAGAGGTTAAAACGAAACCTGTTGTGACATTT


ACAATTCAAAATGGAGAAGTTACGAACCTGAAAGCAGATCCAAATGCTAATAAAAATCAAATCGGGTATCTT


GAAGGAAATGGTAAACATCTTATTACCAACACTCCCAAACGCCCACCAGGTGTTTTTCCTAAAACAGGGGGA


ATTGGTACAATTGTCTATATATTAGTTGGTTCTACTTTTATGATACTTACCATTTGTTCTTTCCGTCGTAAA


CAATTGTAA


 


ORF amino acid sequence (SEQ ID NO: 192):


MKKRQKIWRGLSVTLLILSQIPFGILVQGETQDTNQALGKVIVKKTGDNATPLGKATFVLKNDNDKSETSHE


TVEGSGEATFENIKPGDYTLREETAPIGYKKTDKTWKVKVADNGATIIEGMDADKAEKRKEVLNAQYPKSAI


YEDTKENYPLVNVEGSKVGEQYKALNPINGKDGRREIAEGWLSKKITGVNDLDKNKYKIELTVEGKTTVETK


ELNQPLDVVVLLDNSNSMNNERANNSQRALKAGEAVEKLIDKITSNKDNRVALVTYASTIFDGTEATVSKGV


ADQNGKALNDSVSWDYHKTTFTATTHNYSYLNLTNDANEVNILKSRIPKEAEHINGDRTLYQFGATFTQKAL


MKANEILETQSSNARKKLIFHVTDGVPTMSYAINFNPYISTSYQNQFNSFLNKIPDRSGILQEDFIINGDDY


QIVKGDGESFKLFSDRKVPVTGGTTQAAYRVPQNQLSVMSNEGYAINSGYIYLYWRDYNWVYPFDPKTKKVS


ATKQIKTHGEPTTLYFNGNIRPKGYDIFTVGIGVNGDPGATPLEAEKFMQSISSKTENYTNVDDTNKIYDEL


NKYFKTIVEEKHSIVDGNVTDPMGEMIEFQLKNGQSFTHDDYVLVGNDGSQLKNGVALGGPNSDGGILKDVT


VTYDKTSQTIKINHLNLGSGQKVVLTYDVRLKDNYISNKFYNTNNRTTLSPKSEKEPNTIRDFPIPKIRDVR


EFPVLTISNQKKMGEVEFIKVNKDKHSESLLGAKFQLQIEKDFSGYKQFVPEGSDVTTKNDGKIYFKALQDG


NYKLYEISSPDGYIEVKTKPVVTFTIQNGEVTNLKADPNANKNQIGYLEGNGKHLITNTPKRPPGVFPKTGG


IGTIVYILVGSTFMILTICSFRRKQL


 


Strain IC246


ORF DNA sequence (SEQ ID NO: 212):


ATGAAAAAGAGACAAAAAATATGGAGAGGGTTATCAGTTACTTTACTAATCCTGTCCCAAATTCCATTTGGT


ATATTGGTACAAGGTGAAACCCAAGATACCAATCAAGCACTTGGAAAAGTAATTGTTAAAAAAACGGGAGAC


AATGCTACACCATTAGGCAAAGCGACTTTTGTGTTAAAAAATGACAATGATAAGTCAGAAACAAGTCACGAA


ACGGTAGAGGGTTCTGGAGAAGCAACCTTTGAAAACATAAAACCTGGAGACTACACATTAAGAGAAGAAACA


GCACCAATTGGTTATAAAAAAACTGATAAAACCTGGAAAGTTAAAGTTGCAGATAACGGAGCAACAATAATC


GAGGGTATGGATGCAGATAAAGCAGAGAAACGAAAAGAAGTTTTGAATGCCCAATATCCAAAATCAGCTATT


TATGAGGATACAAAAGAAAATTACCCATTAGTTAATGTAGAGGGTTCCAAAGTTGGTGAACAATACAAAGCA


TTGAATCCAATAAATGGAAAAGATGGTCGAAGAGAGATTGCTGAAGGTTGGTTATCAAAAAAAAATACAGGG


GTCAATGATCTCGATAAGAATAAATATAAAATTGAATTAACTGTTGAGGGTAAAACCACTGTTGAAACGAAA


GAACTTAATCAACCACTAGATGTCGTTGTGCTATTAGATAATTCAAATAGTATGAATAATGAAAGAGCCAAT


AATTCTCAAAGAGCATTAAAAGCTGGGGAAGCAGTTGAAAAGCTGATTGATAAAATTACATCAAATAAAGAC


AATAGAGTAGCTCTTGTGACATATGCCTCAACCATTTTTGATGGTACTGAAGCGACCGTATCAAAGGGAGTT


GCCGATCAAAATGGTAAAGCGCTGAATGATAGTGTATCATGGGATTATCATAAAACTACTTTTACAGCAACT


ACACATAATTACAGTTATTTAAATTTAACAAATGATGCTAACGAAGTTAATATTCTAAAGTCAAGAATTCCA


AAGGAAGCGGAGCATATAAATGGGGATCGCACGCTCTATCAATTTGGTGCGACATTTACTCAAAAAGCTCTA


ATGAAAGCAAATGAAATTTTAGAGACACAAAGTTCTAATGCTAGAAAAAAACTTATTTTTCACGTAACTGAT


GGTGTCCCTACGATGTCTTATGCCATAAATTTTAATCCTTATATATCAACATCTTACCAAAACCAGTTTAAT


TCTTTTTTAAATAAAATACCAGATAGAAGTGGTATTCTCCAAGAGGATTTTATAATCAATGGTGATGATTAT


CAAATAGTAAAAGGAGATGGAGAGAGTTTTAAACTGTTTTCGGATAGAAAAGTTCCTGTTACTGGAGGAACG


ACACAAGCAGCTTATCGAGTACCGCAAAATCAACTCTCTGTAATGAGTAATGAGGGATATGCAATTAATAGT


GGATATATTTATCTCTATTGGAGAGATTACAACTGGGTCTATCCATTTGATCCTAAGACAAAGAAAGTTTCT


GCAACGAAACAAATCAAAACTCATGGTGAGCCAACAACATTATACTTTAATGGAAATATAAGACCTAAAGGT


TATGACATTTTTACTGTTGGGATTGGTGTAAACGGAGATCCTGGTGCAACTCCTCTTGAAGCTGAGAAATTT


ATGCAATCAATATCAAGTAAAACAGAAAATTATACTAATGTTGATGATACAAATAAAATTTATGATGAGCTA


AATAAATACTTTAAAACAATTGTTGAGGAAAAACATTCTATTGTTGATGGAAATGTGACTGATCCTATGGGA


GAGATGATTGAATTCCAATTAAAAAATGGTCAAAGTTTTACACATGATGATTACGTTTTGGTTGGAAATGAT


GGCAGTCAATTAAAAAATGGTGTGGCTCTTGGTGGACCAAACAGTGATGGGGGAATTTTAAAAGATGTTACA


GTGACTTATGATAAGACATCTCAAACCATCAAAATCAATCATTTGAACTTAGGAAGTGGACAAAAAGTAGTT


CTTACCTATGATGTACGTTTAAAAGATAACTATATAAGTAACAAATTTTACAATACAAATAATCGTACAACG


CTAAGTCCGAAGAGTGAAAAAGAACCAAATACTATTCGTGATTTCCCAATTCCCAAAATTCGTGATGTTCGT


GAGTTTCCGGTACTAACCATCAGTAATCAGAAGAAAATGGGTGAGGTTGAATTTATTAAAGTTAATAAAGAC


AAACATTCAGAATCGCTTTTGGGAGCTAAGTTTCAACTTCAGATAGAAAAAGATTTTTCTGGGTATAAGCAA


TTTGTTCCAGAGGGAAGTGATGTTACAACAAAGAATGATGGTAAAATTTATTTTAAAGCACTTCAAGATGGT


AACTATAAATTATATGAAATTTCAAGTCCAGATGGCTATATAGAGGTTAAAACGAAACCTGTTGTGACATTT


ACAATTCAAAATGGAGAAGTTACGAACCTGAAAGCAGATCCAAATGCTAATAAAAATCAAATCGGGTATCTT


GAAGGAAATGGTAAACATCTTATTACCAACACTCCCAAACGCCCACCAGGTGTTTTTCCTAAAACAGGGGGA


ATTGGTACAATTGTCTATATATTAGTTGGTTCTACTTTTATGATACTTACCATTTGTTCTTTCCGTCGTAAA


CAATTGTAA


 


ORF amino acid sequence (SEQ ID NO: 193):


MKKRQKIWRGLSVTLLILSQIPFGILVQGETQDTNQALGKVIVKKTGDNATPLGKATFVLKNDNDKSETSHE


TVEGSGEATFENIKPGDYTLREETAPIGYKKTDKTWKVKVADNGATIIEGMDADKAEKRKEVLNAQYPKSAI


YEDTKENYPLVNVEGSKVGEQYKALNPINGKDGRREIAEGWLSKKNTGVNDLDKNKYKIELTVEGKTTVETK


ELNQPLDVVVLLDNSNSMNNERANNSQRALKAGEAVEKLIDKITSNKDNRVALVTYASTIFDGTEATVSKGV


ADQNGKALNDSVSWDYHKTTFTATTHNYSYLNLTNDANEVNILKSRIPKEAEHINGDRTLYQFGATFTQKAL


MKANEILETQSSNARKKLIFHVTDGVPTMSYAINFNPYISTSYQNQFNSFLNKIPDRSGILQEDFIINGDDY


QIVKGDGESFKLFSDRKVPVTGGTTQAAYRVPQNQLSVMSNEGYAINSGYIYLYWRDYNWVYPFDPKTKKVS


ATKQIKTHGEPTTLYFNGNIRPKGYDIFTVGIGVNGDPGATPLEAEKFMQSISSKTENYTNVDDTNKIYDEL


NKYFKTIVEEKHSIVDGNVTDPMGEMIEFQLKNGQSFTHDDYVLVGNDGSQLKNGVALGGPNSDGGILKDVT


VTYDKTSQTIKINHLNLGSGQKVVLTYDVRLKDNYISNKFYNTNNRTTLSPKSEKEPNTIRDFPIPKIRDVR


EFPVLTISNQKKMGEVEFIKVNKDKHSESLLGAKFQLQIEKDFSGYKQFVPEGSDVTTKNDGKIYFKALQDG


NYKLYEISSPDGYIEVKTKPVVTFTIQNGEVTNLKADPNANKNQIGYLEGNGKHLITNTPKRPPGVFPKTGG


IGTIVYILVGSTFMILTICSFRRKQL


 


Strain IC247


ORF DNA sequence (SEQ ID NO: 213):


ATGAAAAAGAGACAAAAAATATGGAGAGGGTTATCAGTTACTTTACTAATCCTGTCCCAAATTCCATTTGGT


ATATTGGTACAAGGTGAAACCCAAGATACCAATCAAGCACTTGGAAAAGTAATTGTTAAAAAAACGGGAGAC


AATGCTACACCATTAGGCAAAGCGACTTTTGTGTTAAAAAATGACAATGATAAGTCAGAAACAAGTCACGAA


ACGGTAGAGGGTTCTGGAGAAGCAACCTTTGAAAACATAAAACCTGGAGACTACACATTAAGAGAAGAAACA


GCACCAATTGGTTATAAAAAAACTGATAAAACCTGGAAAGTTAAAGTTGCAGATAACGGAGCAACAATAATC


GAGGGTATGGATGCAGATAAAGCAGAGAAACGAAAAGAAGTTTTGAATGCCCAATATCCAAAATCAGCTATT


TATGAGGATACAAAAGAAAATTACCCATTAGTTAATGTAGAGGGTTCCAAAGTTGGTGAACAATACAAAGCA


TTGAATCCAATAAATGGAAAAGATGGTCGAAGAGAGATTGCTGAAGGTTGGTTATCAAAAAAAATTACAGGG


GTCAATGATCTCGATAAGAATAAATATAAAATTGAATTAACTGTTGAGGGTAAAACCACTGTTGAAACGAAA


GAACTTAATCAACCACTAGATGTCGTTGTGCTATTAGATAATTCAAATAGTATGAATAATGAAAGAGCCAAT


AATTCTCAAAGAGCATTAAAAGCTGGGGAAGCAGTTGAAAAGCTGATTGATAAAATTACATCAAATAAAGAC


AATAGAGTAGCTCTTGTGACATATGCCTCAACCATTTTTGATGGTACTGAAGCGACCGTATCAAAGGGAGTT


GCCGATCAAAATGGTAAAGCGCTGAATGATAGTGTATCATGGGATTATCATAAAACTACTTTTACAGCAACT


ACACATAATTACAGTTATTTAAATTTAACAAATGATGCTAACGAAGTTAATATTCTAAAGTCAAGAATTCCA


AAGGAAGCGGAGCATATAAATGGGGATCGCACGCTCTATCAATTTGGTGCGACATTTACTCAAAAAGCTCTA


ATGAAAGCAAATGAAATTTTAGAGACACAAAGTTCTAATGCTAGAAAAAAACTTATTTTTCACGTAACTGAT


GGTGTCCCTACGATGTCTTATGCCATAAATTTTAATCCTTATATATCAACATCTTACCAAAACCAGTTTAAT


TCTTTTTTAAATAAAATACCAGATAGAAGTGGTATTCTCCAAGAGGATTTTATAATCAATGGTGATGATTAT


CAAATAGTAAAAGGAGATGGAGAGAGTTTTAAACTGTTTTCGGATAGAAAAGTTCCTGTTACTGGAGGAACG


ACACAAGCAGCTTATCGAGTACCGCAAAATCAACTCTCTGTAATGAGTAATGAGGGATATGCAATTAATAGT


GGATATATTTATCTCTATTGGAGAGATTACAACTGGGTCTATCCATTTGATCCTAAGACAAAGAAAGTTTCT


GCAACGAAACAAATCAAAACTCATGGTGAGCCAACAACATTATACTTTAATGGAAATATAAGACCTAAAGGT


TATGACATTTTTACTGTTGGGATTGGTGTAAACGGAGATCCTGGTGCAACTCCTCTTGAAGCTGAGAAATTT


ATGCAATCAATATCAAGTAAAACAGAAAATTATACTAATGTTGATGATACAAATAAAATTTATGATGAGCTA


AATAAATACTTTAAAACAATTGTTGAGGAAAAACATTCTATTGTTGATGGAAATGTGACTGATCCTATGGGA


GAGATGATTGAATTCCAATTAAAAAATGGTCAAAGTTTTACACATGATGATTACGTTTTGGTTGGAAATGAT


GGCAGTCAATTAAAAAATGGTGTGGCTCTTGGTGGACCAAACAGTGATGGGGGAATTTTAAAAGATGTTACA


GTGACTTATGATAAGACATCTCAAACCATCAAAATCAATCATTTGAACTTAGGAAGTGGACAAAAAGTAGTT


CTTACCTATGATGTACGTTTAAAAGATAACTATATAAGTAACAAATTTTACAATACAAATAATCGTACAACG


CTAAGTCCGAAGAGTGAAAAAGAACCAAATACTATTCGTGATTTCCCAATTCCCAAAATTCGTGATGTTCGT


GAGTTTCCGGTACTAACCATCAGTAATCAGAAGAAAATGGGTGAGGTTGAATTTATTAAAGTTAATAAAGAC


AAACATTCAGAATCGCTTTTGGGAGCTAAGTTTCAACTTCAGATAGAAAAAGATTTTTCTGGGTATAAGCAA


TTTGTTCCAGAGGGAAGTGATGTTACAACAAAGAATGATGGTAAAATTTATTTTAAAGCACTTCAAGATGGT


AACTATAAATTATATGAAATTTCAAGTCCAGATGGCTATATAGAGGTTAAAACGAAACCTGTTGTGACATTT


ACAATTCAAAATGGAGAAGTTACGAACCTGAAAGCAGATCCAAATGCTAATAAAAATCAAATCGGGTATCTT


GAAGGAAATGGTAAACATCTTATTACCAACACTCCCAAACGCCCACCAGGTGTTTTTCCTAAAACAGGGGGA


ATTGGTACAATTGTCTATATATTAGTTGGTTCTACTTTTATGATACTTACCATTTGTTCTTTCCGTCGTAAA


CAATTGTAA


 


ORF amino acid sequence (SEQ ID NO: 194):


MKKRQKIWRGLSVTLLILSQIPFGILVQGETQDTNQALGKVIVKKTGDNATPLGKATFVLKNDNDKSETSHE


TVEGSGEATFENIKPGDYTLREETAPIGYKKTDKTWKVKVADNGATIIEGMDADKAEKRKEVLNAQYPKSAI


YEDTKENYPLVNVEGSKVGEQYKALNPINGKDGRREIAEGWLSKKITGVNDLDKNKYKIELTVEGKTTVETK


ELNQPLDVVVLLDNSNSMNNERANNSQRALKAGEAVEKLIDKITSNKDNRVALVTYASTIFDGTEATVSKGV


ADQNGKALNDSVSWDYHKTTFTATTHNYSYLNLTNDANEVNILKSRIPKEAEHINGDRTLYQFGATFTQKAL


MKANEILETQSSNARKKLIFHVTDGVPTMSYAINFNPYISTSYQNQFNSFLNKIPDRSGILQEDFIINGDDY


QIVKGDGESFKLFSDRKVPVTGGTTQAAYRVPQNQLSVMSNEGYAINSGYIYLYWRDYNWVYPFDPKTKKVS


ATKQIKTHGEPTTLYFNGNIRPKGYDIFTVGIGVNGDPGATPLEAEKFMQSISSKTENYTNVDDTNKIYDEL


NKYFKTIVEEKHSIVDGNVTDPMGEMIEFQLKNGQSFTHDDYVLVGNDGSQLKNGVALGGPNSDGGILKDVT


VTYDKTSQTIKINHLNLGSGQKVVLTYDVRLKDNYISNKFYNTNNRTTLSPKSEKEPNTIRDFPIPKIRDVR


EFPVLTISNQKKMGEVEFIKVNKDKHSESLLGAKFQLQIEKDFSGYKQFVPEGSDVTTKNDGKIYFKALQDG


NYKLYEISSPDGYIEVKTKPVVTFTIQNGEVTNLKADPNANKNQIGYLEGNGKHLITNTPKRPPGVFPKTGG


IGTIVYILVGSTFMILTICSFRRKQL


 


Strain IC250


ORF DNA sequence (SEQ ID NO: 214):


ATGAGAAAATACCAAAAATTTTCTAAAATATTGACGTTAAGTCTTTTTTGTTTGTCGCAAATACCGCTTAAT


ACCAATGTTTTAGGGGAAAGTACCGTACCGGAAAATGGTGCTAAAGGAAAGTTAGTTGTTAAAAAGACAGAT


GACCAGAACAAACCACTTTCAAAAGCTACCTTTGTTTTAAAAACTACTGCTCAACCAGAAAGTAAAATAGAA


AAAGTAACTGCTGAGCTAACAGGTGAAGCTACTTTTGATAATCTCATACCTGGAGATTATACTTTATCAGAA


GAAACAGCGCCCGAAGGTTATAAAAAGACTAACCAGACTTGGCAAGTTAAGGTTGAGAGTAATGGAAAAACT


ACGATACAAAATAGTGGTGATAAAAATTCCACAATTGGACAAAATCAGGAAGAACTAGATAAGCAGTATCCC


CCCACAGGAATTTATGAAGATACAAAGGAATCTTATAAACTTGAGCATGTTAAAGGTTCAGTTCCAAATGGA


AAGTCAGAGGCAAAAGCAGTTAACCCATATTCAAGTGAAGGTGAGCATATAAGAGAAATTCCAGAGGGAACA


TTATCTAAACGTATTTCAGAAGTAGGTGATTTAGCTCATAATAAATATAAAATTGAGTTAACTGTCAGTGGA


AAAACCATAGTAAAACCAGTGGACAAACAAAAGCCGTTAGATGTTGTCTTCGTACTCGATAATTCTAACTCA


ATGAATAACGATGGCCCAAATTTTCAAAGGCATAATAAAGCCAAGAAAGCTGCCGAAGCTCTTGGGACCGCA


GTAAAAGATATTTTAGGAGCAAACAGTGATAATAGGGTTGCATTAGTTACCTATGGTTCAGATATTTTTGAT


GGTAGGAGTGTAGATGTCGTAAAAGGATTTAAAGAAGATGATAAATATTATGGCCTTCAAACTAAGTTCACA


ATTCAGACAGAGAATTATAGTCATAAACAATTAACAAATAATGCTGAAGAGATTATAAAAAGGATTCCGACA


GAAGCTCCTAAAGCTAAGTGGGGATCTACTACCAATGGATTAACTCCAGAGCAACAAAAGGAGTACTATCTT


AGTAAAGTAGGAGAAACATTTACTATGAAAGCCTTCATGGAGGCAGATGATATTTTGAGTCAAGTAAATCGA


AATAGTCAAAAAATTATTGTTCATGTAACTGATGGTGTTCCTACGAGATCATATGCTATTAATAATTTTAAA


CTGGGTGCATCATATGAAAGCCAATTTGAACAAATGAAAAAAAATGGATATCTAAATAAAAGTAATTTTCTA


CTTACTGATAAGCCCGAGGATATAAAAGGAAATGGGGAGAGTTACTTTTTGTTTCCCTTAGATAGTTATCAA


ACACAGATAATCTCTGGAAACTTACAAAAACTTCATTATTTAGATTTAAATCTTAATTACCCTAAAGGTACA


TTTTATCGAAATGGACCAGTAAGAGAACATGGAACACCAACCAAACTTTATATAAATAGTTTAAAACAGAAA


AATTATGACATCTTTAATTTTGGTATAGATATATCTGGTTTTAGACAAGTTTATAATGAGGATTATAAGAAA


AATCAAGATGGTACTTTTCAAAAATTGAAAGAGGAAGCTTTTGAACTTTCAGATGGGGAAATAACAGAACTA


ATGAAGTCATTCTCTTCTAAACCTGAGTATTATACCCCGATAGTAACTTCATCCGATGCATCTAACAATGAA


ATTTTATCTAAAATTCAGCAACAATTTGAAAAGATTTTAACAAAAGAAAACTCAATTGTTAATGGAACTATA


GAAGATCCTATGGGTGACAAAATCAATTTACAGCTTGGCAACGGACAAACATTGCAACCAAGTGATTATACT


TTACAGGGAAATGATGGAAGTATAATGAAAGATAGCATTGCAACTGGTGGGCCTAATAATGATGGTGGAATA


CTTAAAGGGGTTAAATTAGAATACATCAAAAATAAACTCTACGTTAGAGGTTTGAACTTAGGGGAGGGACAA


AAAGTAACACTCACATATGATGTGAAACTAGATGACAGTTTTATAAGTAACAAATTCTATGACACTAATGGT


AGAACAACATTGAATCCTAAATCAGAGGATCCTAATACACTTAGAGATTTTCCAATCCCTAAAATTCGTGAT


GTGAGAGAATATCCTACAATAACGATTAAAAACGAGAAGAAGTTAGGTGAAATTGAATTTACAAAAGTTGAT


AAAGATAATAATAAGTTGCTTCTCAAAGGAGCTACGTTTGAACTTCAAGAATTTAATGAAGATTATAAACTT


TATTTACCAATAAAAAATAATAATTCAAAAGTAGTGACGGGAGAAAACGGCAAAATTTCTTACAAAGATTTG


AAAGATGGCAAATATCAGTTAATAGAAGCAGTTTCGCCGAAGGATTATCAAAAAATTACTAATAAACCAATT


TTAACTTTTGAAGTTGTTAAAGGATCGATACAAAATATAATAGCTGTTAATAAACAGATTTCTGAATATCAT


GAGGAAGGTGACAAGCATTTAATTACCAACACGCATATTCCACCAAAAGGAATTATTCCGATGACAGGTGGG


AAAGGAATTCTATCTTTCATTTTAATAGGTGGATCTATGATGTCTATTGCAGGTGGAATTTATATTTGGAAA


AGATATAAGAAATCTAGTGATATATCTAGAGAAAAAGATTAA


 


ORF amino acid sequence (SEQ ID NO: 195):


MRKYQKFSKILTLSLFCLSQIPLNTNVLGESTVPENGAKGKLVVKKTDDQNKPLSKATFVLKTTAQPESKIE


KVTAELTGEATFDNLIPGDYTLSEETAPEGYKKTNQTWQVKVESNGKTTIQNSGDKNSTIGQNQEELDKQYP


PTGIYEDTKESYKLEHVKGSVPNGKSEAKAVNPYSSEGEHIREIPEGTLSKRISEVGDLAHNKYKIELTVSG


KTIVKPVDKQKPLDVVFVLDNSNSMNNDGPNFQRHNKAKKAAEALGTAVKDILGANSDNRVALVTYGSDIFD


GRSVDVVKGFKEDDKYYGLQTKFTIQTENYSHKQLTNNAEEIIKRIPTEAPKAKWGSTTNGLTPEQQKEYYL


SKVGETFTMKAFMEADDILSQVNRNSQKIIVHVTDGVPTRSYAINNFKLGASYESQFEQMKKNGYLNKSNFL


LTDKPEDIKGNGESYFLFPLDSYQTQIISGNLQKLHYLDLNLNYPKGTFYRNGPVREHGTPTKLYINSLKQK


NYDIFNFGIDISGFRQVYNEDYKKNQDGTFQKLKEEAFELSDGEITELMKSFSSKPEYYTPIVTSSDASNNE


ILSKIQQQFEKILTKENSIVNGTIEDPMGDKINLQLGNGQTLQPSDYTLQGNDGSIMKDSIATGGPNNDGGI


LKGVKLEYIKNKLYVRGLNLGEGQKVTLTYDVKLDDSFISNKFYDTNGRTTLNPKSEDPNTLRDFPIPKIRD


VREYPTITIKNEKKLGEIEFTKVDKDNNKLLLKGATFELQEFNEDYKLYLPIKNNNSKVVTGENGKISYKDL


KDGKYQLIEAVSPKDYQKITNKPILTFEVVKGSIQNIIAVNKQISEYHEEGDKHLITNTHIPPKGIIPMTGG


KGILSFILIGGSMMSIAGGIYIWKRYKKSSDISREKD


 


Strain IC251


ORF DNA sequence (SEQ ID NO: 215):


ATGAAAAAGAGACAAAAAATATGGAGAGGGTTATCAGTTACTTTACTAATCCTGTCCCAAATTCCATTTGGT


ATATTGGTACAAGGTGAAACCCAAGATACCAATCAAGCACTTGGAAAAGTAATTGTTAAAAAAACGGGAGAC


AATGCTACACCATTAGGCAAAGCGACTTTTGTGTTAAAAAATGACAATGATAAGTCAGAAACAAGTCACGAA


ACGGTAGAGGGTTCTGGAGAAGCAACCTTTGAAAACATAAAACCTGGAGACTACACATTAAGAGAAGAAACA


GCACCAATTGGTTATAAAAAAACTGATAAAACCTGGAAAGTTAAAGTTGCAGATAACGGAGCAACAATAATC


GAGGGTATGGATGCAGATAAAGCAGAGAAACGAAAAGAAGTTTTGAATGCCCAATATCCAAAATCAGCTATT


TATGAGGATACAAAAGAAAATTACCCATTAGTTAATGTAGAGGGTTCCAAAGTTGGTGAACAATACAAAGCA


TTGAATCCAATAAATGGAAAAGATGGTCGAAGAGAGATTGCTGAAGGTTGGTTATCAAAAAAAATTACAGGG


GTCAATGATCTCGATAAGAATAAATATAAAATTGAATTAACTGTTGAGGGTAAAACCACTGTTGAAACGAAA


GAACTTAATCAACCACTAGATGTCGTTGTGCTATTAGATAATTCAAATAGTATGAATAATGAAAGAGCCAAT


AATTCTCAAAGAGCATTAAAAGCTGGGGAAGCAGTTGAAAAGCTGATTGATAAAATTACATCAAATAAAGAC


AATAGAGTAGCTCTTGTGACATATGCCTCAACCATTTTTGATGGTACTGAAGCGACCGTATCAAAGGGAGTT


GCCGATCAAAATGGTAAAGCGCTGAATGATAGTGTATCATGGGATTATCATAAAACTACTTTTACAGCAACT


ACACATAATTACAGTTATTTAAATTTAACAAATGATGCTAACGAAGTTAATATTCTAAAGTCAAGAATTCCA


AAGGAAGCGGAGCATATAAATGGGGATCGCACGCTCTATCAATTTGGTGCGACATTTACTCAAAAAGCTCTA


ATGAAAGCAAATGAAATTTTAGAGACACAAAGTTCTAATGCTAGAAAAAAACTTATTTTTCACGTAACTGAT


GGTGTCCCTACGATGTCTTATGCCATAAATTTTAATCCTTATATATCAACATCTTACCAAAACCAGTTTAAT


TCTTTTTTAAATAAAATACCAGATAGAAGTGGTATTCTCCAAGAGGATTTTATAATCAATGGTGATGATTAT


CAAATAGTAAAAGGAGATGGAGAGAGTTTTAAACTGTTTTCGGATAGAAAAGTTCCTGTTACTGGAGGAACG


ACACAAGCAGCTTATCGAGTACCGCAAAATCAACTCTCTGTAATGAGTAATGAGGGATATGCAATTAATAGT


GGATATATTTATCTCTATTGGAGAGATTACAACTGGGTCTATCCATTTGATCCTAAGACAAAGAAAGTTTCT


GCAACGAAACAAATCAAAACTCATGGTGAGCCAACAACATTATACTTTAATGGAAATATAAGACCTAAAGGT


TATGACATTTTTACTGTTGGGATTGGTGTAAACGGAGATCCTGGTGCAACTCCTCTTGAAGCTGAGAAATTT


ATGCAATCAATATCAAGTAAAACAGAAAATTATACTAATGTTGATGATACAAATAAAATTTATGATGAGCTA


AATAAATACTTTAAAACAATTGTTGAGGAAAAACATTCTATTGTTGATGGAAATGTGACTGATCCTATGGGA


GAGATGATTGAATTCCAATTAAAAAATGGTCAAAGTTTTACACATGATGATTACGTTTTGGTTGGAAATGAT


GGCAGTCAATTAAAAAATGGTGTGGCTCTTGGTGGACCAAACAGTGATGGGGGAATTTTAAAAGATGTTACA


GTGACTTATGATAAGACATCTCAAACCATCAAAATCAATCATTTGAACTTAGGAAGTGGACAAAAAGTAGTT


CTTACCTATGATGTACGTTTAAAAGATAACTATATAAGTAACAAATTTTACAATACAAATAATCGTACAACG


CTAAGTCCGAAGAGTGAAAAAGAACCAAATACTATTCGTGATTTCCCAATTCCCAAAATTCGTGATGTTCGT


GAGTTTCCGGTACTAACCATCAGTAATCAGAAGAAAATGGGTGAGGTTGAATTTATTAAAGTTAATAAAGAC


AAACATTCAGAATCGCTTTTGGGAGCTAAGTTTCAACTTCAGATAGAAAAAGATTTTTCTGGGTATAAGCAA


TTTGTTCCAGAGGGAAGTGATGTTACAACAAAGAATGATGGTAAAATTTATTTTAAAGCACTTCAAGATGGT


AACTATAAATTATATGAAATTTCAAGTCCAGATGGCTATATAGAGGTTAAAACGAAACCTGTTGTGACATTT


ACAATTCAAAATGGAGAAGTTACGAACCTGAAAGCAGATCCAAATGCTAATAAAAATCAAATCGGGTATCTT


GAAGGAAATGGTAAACATCTTATTACCAACACTCCCAAACGCCCACCAGGTGTTTTTCCTAAAACAGGGGGA


ATTGGTACAATTGTCTATATATTAGTTGGTTCTACTTTTATGATACTTACCATTTGTTCTTTCCGTCGTAAA


CAATTGTAA


 


ORF amino acid sequence (SEQ ID NO: 196):


MKKRQKIWRGLSVTLLILSQIPFGILVQGETQDTNQALGKVIVKKTGDNATPLGKATFVLKNDNDKSETSHE


TVEGSGEATFENIKPGDYTLREETAPIGYKKTDKTWKVKVADNGATIIEGMDADKAEKRKEVLNAQYPKSAI


YEDTKENYPLVNVEGSKVGEQYKALNPINGKDGRREIAEGWLSKKITGVNDLDKNKYKIELTVEGKTTVETK


ELNQPLDVVVLLDNSNSMNNERANNSQRALKAGEAVEKLIDKITSNKDNRVALVTYASTIFDGTEATVSKGV


ADQNGKALNDSVSWDYHKTTFTATTHNYSYLNLTNDANEVNILKSRIPKEAEHINGDRTLYQFGATFTQKAL


MKANEILETQSSNARKKLIFHVTDGVPTMSYAINFNPYISTSYQNQFNSFLNKIPDRSGILQEDFIINGDDY


QIVKGDGESFKLFSDRKVPVTGGTTQAAYRVPQNQLSVMSNEGYAINSGYIYLYWRDYNWVYPFDPKTKKVS


ATKQIKTHGEPTTLYFNGNIRPKGYDIFTVGIGVNGDPGATPLEAEKFMQSISSKTENYTNVDDTNKIYDEL


NKYFKTIVEEKHSIVDGNVTDPMGEMIEFQLKNGQSFTHDDYVLVGNDGSQLKNGVALGGPNSDGGILKDVT


VTYDKTSQTIKINHLNLGSGQKVVLTYDVRLKDNYISNKFYNTNNRTTLSPKSEKEPNTIRDFPIPKIRDVR


EFPVLTISNQKKMGEVEFIKVNKDKHSESLLGAKFQLQIEKDFSGYKQFVPEGSDVTTKNDGKIYFKALQDG


NYKLYEISSPDGYIEVKTKPVVTFTIQNGEVTNLKADPNANKNQIGYLEGNGKHLITNTPKRPPGVFPKTGG


IGTIVYILVGSTFMILTICSFRRKQL


 


Strain IC253


ORF DNA sequence (SEQ ID NO: 216):


ATGAGAAAATACCAAAAATTTTCTAAAATATTGACGTTAAGTCTTTTTTGTTTGTCGCAAATACCGCTTAAT


ACCAATGTTTTAGGGGAAAGTACCGTACCGGAAAATGGTGCTAAAGGAAAGTTAGTTGTTAAAAAGACAGAT


GACCAGAACAAACCACTTTCAAAAGCTACCTTTGTTTTAAAAACTACTGCTCATCCAGAAAGTAAAATAGAA


AAAGTAACTGCTGAGCTAACAGGTGAAGCTACTTTTGATAATCTCATACCTGGAGATTATACTTTATCAGAA


GAAACAGCGCCCGAAGGTTATAAAAAGACTAACCAGACTTGGCAAGTTAAGGTTGAGAGTAATGGAAAAACT


ACGATACAAAATAGTGGTGATAAAAATTCCACAATTGGACAAAATCACGAAGAACTAGATAAGCAGTATCCC


CCCACAGGAATTTATGAAGATACAAAGGAATCTTATAAACTTGAGCATGTTAAAGGTTCAGTTCCAAATGGA


AAGTCAGAGGCAAAAGCAGTTAACCCATATTCAAGTGAAGGTGAGCATATAAGAGAAATTCCAGAGGGAACA


TTATCTAAACGTATTTCAGAAGTAGGTGATTTAGCTCATAATAAATATAAAATTGAGTTAACTGTCAGTGGA


AAAACCATAGTAAAACCAGTGGACAAACAAAAGCCGTTAGATGTTGTCTTCGTACTCGATAATTCTAACTCA


ATGAATAACGATGGCCCAAATTTTCAAAGGCATAATAAAGCCAAGAAAGCTGCCGAAGCTCTTGGGACCGCA


GTAAAAGATATTTTAGGAGCAAACAGTGATAATAGGGTTGCATTAGTTACCTATGGTTCAGATATTTTTGAT


GGTAGGAGTGTAGATGTCGTAAAAGGATTTAAAGAAGATGATAAATATTATGGCCTTCAAACTAAGTTCACA


ATTCAGACAGAGAATTATAGTCATAAACAATTAACAAATAATGCTGAAGAGATTATAAAAAGGATTCCTACA


GAAGCTCCTAGAGCTAAATGGGGATCAACTACAAACGGACTTACTCCAGAGCAACAAAAGCAGTACTATCTT


AGTAAAGTAGGGGAAACATTTACTATGAAAGCCTTCATGGAGGCAGATGATATTTTGAGTCAAGTAGATCGA


AATAGTCAAAAAATTATTGTTCATATAACTGATGGTGTTCCAACAAGATCATATGCTATTAATAATTTTAAA


TTGGGTGCATCATATGAAAGCCAATTTGAACAAATGAAAAAAAATGGATATCTAAATAAAAGTAATTTTCTA


CTTACTGATAAGCCCGAGGATATAAAAGGAAATGGGGAGAGTTACTTTTTGTTTCCCTTAGATAGTTATCAA


ACACAGATAATCTCTGGAAACTTACAAAAACTTCATTATTTAGATTTAAATCTTAATTACCCTAAAGGTACA


ATTTATCGAAATGGACCAGTAAGAGAACATGGAACACCAACCAAACTTTATATAAATAGTTTAAAACAGAAA


AATTATGACATCTTTAATTTTGGTATAGATATATCTGCTTTTAGACAAGTTTATAATGAGGATTATAAGAAA


AATCAAGATGGTACTTTTCAAAAATTGAAAGAGGAAGCTTTTGAACTTTCAGATGGGGAAATAACAGAACTA


ATGAAGTCATTCTCTTCTAAACCTGAGTATTATACCCCGATAGTAACTTCATCCGATGCATCTAACAATGAA


ATTTTATCTAAAATTCAGCAACAATTTGAAAAGGTTTTAACAAAAGAAAACTCAATTGTTAATGGAACTATA


GAAGATCCTATGGGTGACAAAATCAATTTACAGCTTGGCAACGGACAAACATTGCAACCAAGTGATTATACT


TTACAGGGAAATGATGGAAGTATAATGAAAGATAGCATTGCAACTGGTGGGCCTAATAATGATGGTGGAATA


CTTAAAGGGGTTAAATTAGAATACATCAAAAATAAACTCTACGTTAGAGGTTTGAACTTAGGGGAGGGACAA


AAAGTAACACTCACATATGATGTGAAACTAGATGACAGTTTTATAAGTAACAAATTCTATGACACTAATGGT


AGAACAACATTGAATCCTAAATCAGAGGATCCTAATACACTTAGAGATTTTCCAATCCCTAAAATTCGTGAT


GTGAGAGAATATCCTACAATAACGATTAAAAACGAGAAGAAGTTAGGTGAAATTGAATTTACAAAAGTTGAT


AAAGATAATAATAAGTTGCTTCTCAAAGGAGCTACGTTTGAACTTCAAGAATTTAATGAAGATTATAAACTT


TATTTACCAATAAAAAATAATAATTCAAAAGTAGTGACGGGAGAAAACGGCAAAATTTCTTACAAAGATTTG


AAAGATGGCAAATATCAGTTAATAGAAGCAGTTTCGCCGAAGGATTATCAAAAAATTACTAATAAACCAATT


TTAACTTTTGAAGTTGTTAAAGGATCGATACAAAATATAATAGCTGTTAATAAACAGATTTCTGAATATCAT


GAGGAAGGTGACAAGCATTTAATTACCAACACGCATATTCCACCAAAAGGAATTATTCCGATGACAGGTGGG


AAAGGAATTCTATCTTTCATTTTAATAGGTGGATCTATGATGTCTATTGCAGGTGGAATTTATATTTGGAAA


AGATATAAGAAATCTAGTGATATATCTAGAGAAAAAGATTAA


 


ORF amino acid sequence (SEQ ID NO: 197):


MRKYQKFSKILTLSLFCLSQIPLNTNVLGESTVPENGAKGKLVVKKTDDQNKPLSKATFVLKTTAHPESKIE


KVTAELTGEATFDNLIPGDYTLSEETAPEGYKKTNQTWQVKVESNGKTTIQNSGDKNSTIGQNHEELDKQYP


PTGIYEDTKESYKLEHVKGSVPNGKSEAKAVNPYSSEGEHIREIPEGTLSKRISEVGDLAHNKYKIELTVSG


KTIVKPVDKQKPLDVVFVLDNSNSMNNDGPNFQRHNKAKKAAEALGTAVKDILGANSDNRVALVTYGSDIFD


GRSVDVVKGFKEDDKYYGLQTKFTIQTENYSHKQLTNNAEEIIKRIPTEAPRAKWGSTTNGLTPEQQKQYYL


SKVGETFTMKAFMEADDILSQVDRNSQKIIVHITDGVPTRSYAINNFKLGASYESQFEQMKKNGYLNKSNFL


LTDKPEDIKGNGESYFLFPLDSYQTQIISGNLQKLHYLDLNLNYPKGTIYRNGPVREHGTPTKLYINSLKQK


NYDIFNFGIDISAFRQVYNEDYKKNQDGTFQKLKEEAFELSDGEITELMKSFSSKPEYYTPIVTSSDASNNE


ILSKIQQQFEKVLTKENSIVNGTIEDPMGDKINLQLGNGQTLQPSDYTLQGNDGSIMKDSIATGGPNNDGGI


LKGVKLEYIKNKLYVRGLNLGEGQKVTLTYDVKLDDSFISNKFYDTNGRTTLNPKSEDPNTLRDFPIPKIRD


VREYPTITIKNEKKLGEIEFTKVDKDNNKLLLKGATFELQEFNEDYKLYLPIKNNNSKVVTGENGKISYKDL


KDGKYQLIEAVSPKDYQKITNKPILTFEVVKGSIQNIIAVNKQISEYHEEGDKHLITNTHIPPKGIIPMTGG


KGILSFILIGGSMMSIAGGIYIWKRYKKSSDISREKD


 


Strain IC289


ORF DNA sequence (SEQ ID NO: 217):


ATGAGAAAATACCAAAAATTTTCTAAAATATTGACGTTAAGTCTTTTTTGTTTGTCGCAAATACCGCTTAAT


ACCAATGTTTTAGGGGAAAGTACCGTACCGGAAAATGGTGCTAAAGGAAAGTTAGTTGTTAAAAAGACAGAT


GACCAGAACAAACCACTTTCAAAAGCTACCTTTGTTTTAAAACCTACTTCACACTCAGAAAGCAAAGTAGAA


AAAGTAACTACTGAGGTAACAGGTGAAGCTACTTTTGATAATCTCACACCTGGAGATTACACTTTATCAGAA


GAAACGGCACCCGAAGGATACAAAAAGACTACCCAGACTTGGCAAGTTAAGGTTGAGAGTAATGGAAAAACT


ACGATACAAAATAGTGATGATAAAAAATCTATAATTGAACAAAGGCAAGAGGAACTAGATAAGCAGTATCCC


CTTACAGGAGCTTATGAAGATACAAAAGAATCTTATAATCTTGAGCATGTTAAAAATTCAATTCCAAATGGG


AAATTAGAGGCAAAAGCAGTTAATCCATATTCAAGTGAAGGTGAGCACATAAGAGAAATTCAAGAGGGAACA


TTATCTAAACGTATTTCAGAAGTAAATGATTTGGATCATAATAAATATAAAATTGAGTTAACTGTTAGCGGT


AAATCCATAATAAAAACTATAAATAAAGATGAACCTCTGGATGTTGTTTTTGTTCTTGATAATTCAAATTCT


ATGAAGAATAATGGAAAAAATAACAAGGCAAAAAAGGCAGGTGAAGCAGTAGAAACAATTATAAAAGATGTT


TTAGGAGCAAATGTTGAAAACCGAGCAGCTTTAGTTACTTATGGTTCAGATATTTTTGATGGAAGGACAGTT


AAAGTTATAAAAGGTTTTAAAGAGGATCCTTATTATGGACTTGAAACTAGTTTCACAGTTCAGACAAATGAT


TATAGCTATAAAAAGTTCACTAATATTGCTGCTGATATTATAAAAAAGATCCCTAAAGAAGCTCCAGAAGCT


AAGTGGGGGGGGACAAGTCTAGGATTAACTCCAGAAAAAAAGAGGGAATATGATTTAAGTAAAGTAGGTGAG


ACCTTTACAATGAAAGCTTTTATGGAGGCAGATACCTTGTTAAGTAGTATACAGCGTAAGAGTAGAAAGATT


ATTGTTCATCTAACTGACGGTGTTCCAACAAGATCATATGCCATTAATAGTTTTGTAAAAGGTTCAACATAC


GCAAATCAATTTGAGAGAATAAAAGAAAAAGGTTATTTAGACAAAAATAATTATTTTATAACTGATGATCCA


GAAAAGATCAAAGGCAATGGGGAGAGTTACTTTTTGTTTCCCTTAGATAGTTATCAAACACAGATAATTTCT


GGAAACTTACAAAAACTTCATTATTTAGATTTAAATCTTAATTACCCTAAAGGTACAATTTATAGAAATGGA


CCAGTAAGAGAACATGGAACACCAACCAAACTTTATATAAATAGTTTAAAACAGAAAAATTATGACATCTTT


AATTTTGGTATAGATATATCTGGTTTTAGACAAGTTTATAATGAGGATTATAAGAAAAATCAAGATGGTACT


TTTCAAAAATTGAAGGAGGAAGCTTTTGAACTTTCAGATGGGGAAATAACAGAACTAATGAATTCATTCTCT


TCTAAACCTGAGTATTATACCCCGATAGTAACTTCAGCTGATGTATCTAATAATGAAATTTTATCTAAAATT


CAGCAACAATTTGAAAAGATTTTAACAAAGGAAAACTCAATTGTTAATGGAACTATAGAAGATCCTATGGGT


GATAAAATCAATTTACATCTTGGCAACGGACAAACATTGCAACCAAGTGATTATACTTTACAGGGAAATGAT


GGAAGTATAATGAAAGATAGCATTGCAACTGGAGGGCCTAATAATGATGGTGGGATACTTAAAGGGGTTAAA


TTAGAATACATCAAAAATAAACTCTACGTTAGAGGTTTGAACTTAGGGGAGGGGCAAAAAGTAACACTCACA


TATGATGTGAAACTAGATGACAGTTTTATTAGTAACAAATTCTATGACACTAATGGTAGAACAACATTGAAT


CCTAAATCAGAGGAACCCGATACACTTAGAGATTTTCCAATCCCTAAAATTCGTGATGTGAGAGAATATCCT


ACAATAACGATTAAAAACGAGAAGAAGTTAGGTGAAATTGAATTTACAAAAGTTGATAAAGATAATAATAAG


TTGCTTCTCAAAGGAGCTACATTTGAACTTCAAGAATTTAATGAAGATTATAAACTTTATTTACCAATAAAA


AATAATAATTCAAAAGTAGTGACGGGAGAAAACGGCAAAATTTCTTACAAAGATTTGAAAGATGGCAAATAT


CAGTTAATAGAAGCAGTTTCGCCGAAGGATTATCAAAAAATTACTAATAAACCAATTTTAACTTTTGAAGTT


GTTAAAGGATCGATACAAAATATAATAGCTGTTAATAAACAGATTTCTGAATATCATGAGGAAGGTGACAAG


CATTTAATTACCAACACGCATATTCCACCAAAAGGAATTATTCCGATGACAGGTGGGAAAGGAATTCTATCT


TTCATTTTAATAGGTGGAGCTATGATGTCTATTGCAGGTGGAATTTATATTTGGAAAAGACATAAGAAATCT


AGTGATGCATCAATCGAGAAAGATTAA


 


ORF amino acid sequence (SEQ ID NO: 198):


MRKYQKFSKILTLSLFCLSQIPLNTNVLGESTVPENGAKGKLVVKKTDDQNKPLSKATFVLKPTSHSESKVE


KVTTEVTGEATFDNLTPGDYTLSEETAPEGYKKTTQTWQVKVESNGKTTIQNSDDKKSIIEQRQEELDKQYP


LTGAYEDTKESYNLEHVKNSIPNGKLEAKAVNPYSSEGEHIREIQEGTLSKRISEVNDLDHNKYKIELTVSG


KSIIKTINKDEPLDVVFVLDNSNSMKNNGKNNKAKKAGEAVETIIKDVLGANVENRAALVTYGSDIFDGRTV


KVIKGFKEDPYYGLETSFTVQTNDYSYKKFTNIAADIIKKIPKEAPEAKWGGTSLGLTPEKKREYDLSKVGE


TFTMKAFMEADTLLSSIQRKSRKIIVHLTDGVPTRSYAINSFVKGSTYANQFERIKEKGYLDKNNYFITDDP


EKIKGNGESYFLFPLDSYQTQIISGNLQKLHYLDLNLNYPKGTIYRNGPVREHGTPTKLYINSLKQKNYDIF


NFGIDISGFRQVYNEDYKKNQDGTFQKLKEEAFELSDGEITELMNSFSSKPEYYTPIVTSADVSNNEILSKI


QQQFEKILTKENSIVNGTIEDPMGDKINLHLGNGQTLQPSDYTLQGNDGSIMKDSIATGGPNNDGGILKGVK


LEYIKNKLYVRGLNLGEGQKVTLTYDVKLDDSFISNKFYDTNGRTTLNPKSEEPDTLRDFPIPKIRDVREYP


TITIKNEKKLGEIEFTKVDKDNNKLLLKGATFELQEFNEDYKLYLPIKNNNSKVVTGENGKISYKDLKDGKY


QLIEAVSPKDYQKITNKPILTFEVVKGSIQNIIAVNKQISEYHEEGDKHLITNTHIPPKGIIPMTGGKGILS


FILIGGAMMSIAGGIYIWKRHKKSSDASIEKD


 


Strain IC291


ORF DNA sequence (SEQ ID NO: 218):


ATGAGAAAATACCAAAAATTTTCTAAAATATTGACGTTAAGTCTTTTTTGTTTGTCGCAAATACCGCTTAAT


ACCAATGTTTTAGGGGAAAGTACCGTACCGGAAAATGGTGCTAAAGGAAAGTTAGTTGTTAAAAAGACAGAT


GACCAGAACAAACCACTTTCAAAAGCTACCTTTGTTTTAAAAACTACTGCTCATCCAGAAAGTAAAATAGAA


AAAGTAACTGCTGAGCTAACAGGTGAAGCTACTTTTGATAATCTCATACCTGGAGATTATACTTTATCAGAA


GAAACAGCGCCCGAAGGTTATAAAAAGACTAACCAGACTTGGCAAGTTAAGGTTGAGAGTAATGGAAAAACT


ACGATACAAAATAGTGGTGATAAAAATTCCACAATTGGACAAAATCAGGAAGAACTAGATAAGCAGTATCCC


CCCACAGGAATTTATGAAGATACAAAGGAATCTTATAAACTTGAGCATGTTAAAGGTTCAGTTCCAAATGGA


AAGTCAGAGGCAAAAGCAGTTAACCCATATTCAAGTGAAGGTGAGCATATAAGAGAAATTCCAGAGGGAACA


TTATCTAAACGTATTTCAGAAGTAGGTGATTTAGCTCATAATAAATATAAAATTGAGTTAACTGTCAGTGGA


AAAACCATAGTAAAACCAGTGGACAAACAAAAGCCGTTAGATGTTGTCTTCGTACTCGATAATTCTAACTCA


ATGAATAACGATGGCCCAAATTTTCAAAGGCATAATAAAGCCAAGAAAGCTGCCGAAGCTCTTGGGACCGCA


GTAAAAGATATTTTAGGAGCAAACAGTGATAATAGGGTTGCATTAGTTACCTATGGTTCAGATATTTTTGAT


GGTAGGAGTGTAGATGTCGTAAAAGGATTTAAAGAAGATGATAAATATTATGGCCTTCAAACTAAGTTCACA


ATTCAGACAGAGAATTATAGTCATAAACAATTAACAAATAATGCTGAAGAGATTATAAAAAGGATTCCGACA


GAAGCTCCTAAAGCTAAGTGGGGATCTACTACCAATGGATTAACTCCAGAGCAACAAAAGGAGTACTATCTT


AGTAAAGTAGGAGAAACATTTACTATGAAAGCCTTCATGGAGGCAGATGATATTTTGAGTCAAGTAAATCGA


AATAGTCAAAAAATTATTGTTCATGTAACTGATGGTGTTCCTACGAGATCATATGCTATTAATAATTTTAAA


CTGGGTGCATCATATGAAAGCCAATTTGAACAAATGAAAAAAAATGGATATCTAAATAAAAGTAATTTTCTA


CTTACTGATAAGCCCGAGGATATAAAAGGAAATGGGGAGAGTTACTTTTTGTTTCCCTTAGATAGTTATCAA


ACACAGATAATCTCTGGAAACTTACAAAAACTTCATTATTTAGATTTAAATCTTAATTACCCTAAAGGTACA


TTTTATCGAAATGGACCAGTAAGAGAACATGGAACACCAACCAAACTTTATATAAATAGTTTAAAACAGAAA


AATTATGACATCTTTAATTTTGGTATAGATATATCTGGTTTTAGACAAGTTTATAATGAGGATTATAAGAAA


AATCAAGATGGTACTTTTCAAAAATTGAAAGAGGAAGCTTTTGAACTTTCAGATGGGGAAATAACAGAACTA


ATGAAGTCATTCTCTTCTAAACCTGAGTATTATACCCCGATAGTAACTTCATCCGATGCATCTAACAATGAA


ATTTTATCTAAAATTCAGCAACAATTTGAAAAGATTTTAACAAAAGAAAACTCAATTGTTAATGGAACTATA


GAAGATCCTATGGGTGACAAAATCAATTTACAGCTTGGCAACGGACAAACATTGCAACCAAGTGATTATACT


TTACAGGGAAATGATGGAAGTATAATGAAAGATAGCATTGCAACTGGTGGGCCTAATAATGATGGTGGAATA


CTTAAAGGGGTTAAATTAGAATACATCAAAAATAAACTCTACGTTAGAGGTTTGAACTTAGGGGAGGGACAA


AAAGTAACACTCACATATGATGTGAAACTAGATGACAGTTTTATAAGTAACAAATTCTATGACACTAATGGT


AGAACAACATTGAATCCTAAATCAGAGGATCCTAATACACTTAGAGATTTTCCAATCCCTAAAATTCGTGAT


GTGAGAGAATATCCTACAATAACGATTAAAAACGAGAAGAAGTTAGGTGAAATTGAATTTACAAAAGTTGAT


AAAGATAATAATAAGTTGCTTCTCAAAGGAGCTACGTTTGAACTTCAAGAATTTAATGAAGATTATAAACTT


TATTTACCAATAAAAAATAATAATTCAAAAGTAGTGACGGGAGAAAACGGCAAAATTTCTTACAAAGATTTG


AAAGATGGCAAATATCAGTTAATAGAAGCAGTTTCGCCGAAGGATTATCAAAAAATTACTAATAAACCAATT


TTAACTTTTGAAGTTGTTAAAGGATCGATACAAAATATAATAGCTGTTAATAAACAGATTTCTGAATATCAT


GAGGAAGGTGACAAGCATTTAATTACCAACACGCATATTCCACCAAAAGGAATTATTCCGATGACAGGTGGG


AAAGGAATTCTATCTTTCATTTTAATAGGTGGATCTATGATGTCTATTGCAGGTGGAATTTATATTTGGAAA


AGATATAAGAAATCTAGTGATATATCTAGAGAAAAAGATTAA


 


ORF amino acid sequence (SEQ ID NO: 199):


MRKYQKFSKILTLSLFCLSQIPLNTNVLGESTVPENGAKGKLVVKKTDDQNKPLSKATFVLKTTAHPESKIE


KVTAELTGEATFDNLIPGDYTLSEETAPEGYKKTNQTWQVKVESNGKTTIQNSGDKNSTIGQNQEELDKQYP


PTGIYEDTKESYKLEHVKGSVPNGKSEAKAVNPYSSEGEHIREIPEGTLSKRISEVGDLAHNKYKIELTVSG


KTIVKPVDKQKPLDVVFVLDNSNSMNNDGPNFQRHNKAKKAAEALGTAVKDILGANSDNRVALVTYGSDIFD


GRSVDVVKGFKEDDKYYGLQTKFTIQTENYSHKQLTNNAEEIIKRIPTEAPKAKWGSTTNGLTPEQQKEYYL


SKVGETFTMKAFMEADDILSQVNRNSQKIIVHVTDGVPTRSYAINNFKLGASYESQFEQMKKNGYLNKSNFL


LTDKPEDIKGNGESYFLFPLDSYQTQIISGNLQKLHYLDLNLNYPKGTFYRNGPVREHGTPTKLYINSLKQK


NYDIFNFGIDISGFRQVYNEDYKKNQDGTFQKLKEEAFELSDGEITELMKSFSSKPEYYTPIVTSSDASNNE


ILSKIQQQFEKILTKENSIVNGTIEDPMGDKINLQLGNGQTLQPSDYTLQGNDGSIMKDSIATGGPNNDGGI


LKGVKLEYIKNKLYVRGLNLGEGQKVTLTYDVKLDDSFISNKFYDTNGRTTLNPKSEDPNTLRDFPIPKIRD


VREYPTITIKNEKKLGEIEFTKVDKDNNKLLLKGATFELQEFNEDYKLYLPIKNNNSKVVTGENGKISYKDL


KDGKYQLIEAVSPKDYQKITNKPILTFEVVKGSIQNIIAVNKQISEYHEEGDKHLITNTHIPPKGIIPMTGG


KGILSFILIGGSMMSIAGGIYIWKRYKKSSDISREKD


 


Strain IC304


ORF DNA sequence (SEQ ID NO: 219):


ATGAGAAAATACCAAAAATTTTCTAAAATATTGACGTTAAGTCTTTTTTGTTTGTCGCAAATACCGCTTAAT


ACCAATGTTTTAGGGGAAAGTACCGTACCGGAAAATGGTGCTAAAGGAAAGTTAGTTGTTAAAAAGACAGAT


GACCAGAACAAACCACTTTCAAAAGCTACCTTTGTTTTAAAAACTACTGCTCAACCAGAAAGTAAAATAGAA


AAAGTAACTGCTGAGCTAACAGGTGAAGCTACTTTTGATAATCTCATACCTGGAGATTATACTTTATCAGAA


GAAACAGCGCCCGAAGGTTATAAAAAGACTAACCAGACTTGGCAAGTTAAGGTTGAGAGTAATGGAAAAACT


ACGATACAAAATAGTGGTGATAAAAATTCCACAATTGGACAAAATCAGGAAGAACTAGATAAGCAGTATCCC


CCCACAGGAATTTATGAAGATACAAAGGAATCTTATAAACTTGAGCATGTTAAAGGTTCAGTTCCAAATGGA


AAGTCAGAGGCAAAAGCAGTTAACCCATATTCAAGTGAAGGTGAGCATATAAGAGAAATTCCAGAGGGAACA


TTATCTAAACGTATTTCAGAAGTAGGTGATTTAGCTCATAATAAATATAAAATTGAGTTAACTGTCAGTGGA


AAAACCATAGTAAAACCAGTGGACAAACAAAAGCCGTTAGATGTTGTCTTCGTACTCGATAATTCTAACTCA


ATGAATAACGATGGCCCAAATTTTCAAAGGCATAATAAAGCCAAGAAAGCTGCCGAAGCTCTTGGGACCGCA


GTAAAAGATATTTTAGGAGCAAACAGTGATAATAGGGTTGCATTAGTTACCTATGGTTCAGATATTTTTGAT


GGTAGGAGTGTAGATGTCGTAAAAGGATTTAAAGAAGATGATAAATATTATGGCCTTCAAACTAAGTTCACA


ATTCAGACAGAGAATTATAGTCATAAACAATTAACAAATAATGCTGAAGAGATTATAAAAAGGATTCCGACA


GAAGCTCCTAAAGCTAAGTGGGGATCTACTACCAATGGATTAACTCCAGAGCAACAAAAGGAGTACTATCTT


AGTAAAGTAGGAGAAACATTTACTATGAAAGCCTTCATGGAGGCAGATGATATTTTGAGTCAAGTAAATCGA


AATAGTCAAAAAATTATTGTTCATGTAACTGATGGTGTTCCTACGAGATCATATGCTATTAATAATTTTAAA


CTGGGTGCATCATATGAAAGCCAATTTGAACAAATGAAAAAAAATGGATATCTAAATAAAAGTAATTTTCTA


CTTACTGATAAGCCCGAGGATATAAAAGGAAATGGGGAGAGTTACTTTTTGTTTCCCTTAGATAGTTATCAA


ACACAGATAATCTCTGGAAACTTACAAAAACTTCATTATTTAGATTTAAATCTTAATTACCCTAAAGGTACA


TTTTATCGAAATGGACCAGTAAGAGAACATGGAACACCAACCAAACTTTATATAAATAGTTTAAAACAGAAA


AATTATGACATCTTTAATTTTGGTATAGATATATCTGGTTTTAGACAAGTTTATAATGAGGATTATAAGAAA


AATCAAGATGGTACTTTTCAAAAATTGAAAGAGGAAGCTTTTGAACTTTCAGATGGGGAAATAACAGAACTA


ATGAAGTCATTCTCTTCTAAACCTGAGTATTATACCCCGATAGTAACTTCATCCGATGCATCTAACAATGAA


ATTTTATCTAAAATTCAGCAACAATTTGAAAAGATTTTAACAAAAGAAAACTCAATTGTTAATGGAACTATA


GAAGATCCTATGGGTGACAAAATCAATTTACAGCTTGGCAACGGACAAACATTGCAACCAAGTGATTATACT


TTACAGGGAAATGATGGAAGTATAATGAAAGATAGCATTGCAACTGGTGGGCCTAATAATGATGGTGGAATA


CTTAAAGGGGTTAAATTAGAATACATCAAAAATAAACTCTACGTTAGAGGTTTGAACTTAGGGGAGGGACAA


AAAGTAACACTCACATATGATGTGAAACTAGATGACAGTTTTATAAGTAACAAATTCTATGACACTAATGGT


AGAACAACATTGAATCCTAAATCAGAGGATCCTAATACACTTAGAGATTTTCCAATCCCTAAAATTCGTGAT


GTGAGAGAATATCCTACAATAACGATTAAAAACGAGAAGAAGTTAGGTGAAATTGAATTTACAAAAGTTGAT


AAAGATAATAATAAGTTGCTTCTCAAAGGAGCTACGTTTGAACTTCAAGAATTTAATGAAGATTATAAACTT


TATTTACCAATAAAAAATAATAATTCAAAAGTAGTGACGGGAGAAAACGGCAAAATTTCTTACAAAGATTTG


AAAGATGGCAAATATCAGTTAATAGAAGCAGTTTCGCCGAAGGATTATCAAAAAATTACTAATAAACCAATT


TTAACTTTTGAAGTTGTTAAAGGATCGATACAAAATATAATAGCTGTTAATAAACAGATTTCTGAATATCAT


GAGGAAGGTGACAAGCATTTAATTACCAACACGCATATTCCACCAAAAGGAATTATTCCGATGACAGGTGGG


AAAGGAATTCTATCTTTCATTTTAATAGGTGGATCTATGATGTCTATTGCAGGTGGAATTTATATTTGGAAA


AGATATAAGAAATCTAGTGATATATCTAGAGAAAAAGATTAA


 


ORF amino acid sequence (SEQ ID NO: 200):


MRKYQKFSKILTLSLFCLSQIPLNTNVLGESTVPENGAKGKLVVKKTDDQNKPLSKATFVLKTTAQPESKIE


KVTAELTGEATFDNLIPGDYTLSEETAPEGYKKTNQTWQVKVESNGKTTIQNSGDKNSTIGQNQEELDKQYP


PTGIYEDTKESYKLEHVKGSVPNGKSEAKAVNPYSSEGEHIREIPEGTLSKRISEVGDLAHNKYKIELTVSG


KTIVKPVDKQKPLDVVFVLDNSNSMNNDGPNFQRHNKAKKAAEALGTAVKDILGANSDNRVALVTYGSDIFD


GRSVDVVKGFKEDDKYYGLQTKFTIQTENYSHKQLTNNAEEIIKRIPTEAPKAKWGSTTNGLTPEQQKEYYL


SKVGETFTMKAFMEADDILSQVNRNSQKIIVHVTDGVPTRSYAINNFKLGASYESQFEQMKKNGYLNKSNFL


LTDKPEDIKGNGESYFLFPLDSYQTQIISGNLQKLHYLDLNLNYPKGTFYRNGPVREHGTPTKLYINSLKQK


NYDIFNFGIDISGFRQVYNEDYKKNQDGTFQKLKEEAFELSDGEITELMKSFSSKPEYYTPIVTSSDASNNE


ILSKIQQQFEKILTKENSIVNGTIEDPMGDKINLQLGNGQTLQPSDYTLQGNDGSIMKDSIATGGPNNDGGI


LKGVKLEYIKNKLYVRGLNLGEGQKVTLTYDVKLDDSFISNKFYDTNGRTTLNPKSEDPNTLRDFPIPKIRD


VREYPTITIKNEKKLGEIEFTKVDKDNNKLLLKGATFELQEFNEDYKLYLPIKNNNSKVVTGENGKISYKDL


KDGKYQLIEAVSPKDYQKITNKPILTFEVVKGSIQNIIAVNKQISEYHEEGDKHLITNTHIPPKGIIPMTGG


KGILSFILIGGSMMSIAGGIYIWKRYKKSSDISREKD


 


Strain IC305


ORF DNA sequence (SEQ ID NO: 220):


ATGAGAAAATACCAAAAATTTTCTAAAATATTGACGTTAAGTCTTTTTTGTTTGTCGCAAATACCGCTTAAT


ACCAATGTTTTAGGGGAAAGTACCGTACCGGAAAATGGTGCTAAAGGAAAGTTAGTTGTTAAAAAGACAGAT


GACCAGAACAAACCACTTTCAAAAGCTACCTTTGTTTTAAAAACTACTGCTCATCCAGAAAGTAAAATAGAA


AAAGTAACTGCTGAGCTAACAGGTGAAGCTACTTTTGATAATCTCATACCTGGAGATTATACTTTATCAGAA


GAAACAGCGCCCGAAGGTTATAAAAAGACTAACCAGACTTGGCAAGTTAAGGTTGAGAGTAATGGAAAAACT


ACGATACAAAATAGTGGTGATAAAAATTCCACAATTGGACAAAATCAGGAAGAACTAGATAAGCAGTATCCC


CCCACAGGAATTTATGAAGATACAAAGGAATCTTATAAACTTGAGCATGTTAAAGGTTCAGTTCCAAATGGA


AAGTCAGAGGCAAAAGCAGTTAACCCATATTCAAGTGAAGGTGAGCATATAAGAGAAATTCCAGAGGGAACA


TTATCTAAACGTATTTCAGAAGTAGGTGATTTAGCTCATAATAAATATAAAATTGAGTTAACTGTCAGTGGA


AAAACCATAGTAAAACCAGTGGACAAACAAAAGCCGTTAGATGTTGTCTTCGTACTCGATAATTCTAACTCA


ATGAATAACGATGGCCCAAATTTTCAAAGGCATAATAAAGCCAAGAAAGCTGCCGAAGCTCTTGGGACCGCA


GTAAAAGATATTTTAGGAGCAAACAGTGATAATAGGGTTGCATTAGTTACCTATGGTTCAGATATTTTTGAT


GGTAGGAGTGTAGATGTCGTAAAAGGATTTAAAGAAGATGATAAATATTATGGCCTTCAAACTAAGTTCACA


ATTCAGACAGAGAATTATAGTCATAAACAATTAACAAATAATGCTGAAGAGATTATAAAAAGGATTCCGACA


GAAGCTCCTAAAGCTAAGTGGGGATCTACTACCAATGGATTAACTCCAGAGCAACAAAAGGAGTACTATCTT


AGTAAAGTAGGAGAAACATTTACTATGAAAGCCTTCATGGAGGCAGATGATATTTTGAGTCAAGTAAATCGA


AATAGTCAAAAAATTATTGTTCATGTAACTGATGGTGTTCCTACGAGATCATATGCTATTAATAATTTTAAA


CTGGGTGCATCATATGAAAGCCAATTTGAACAAATGAAAAAAAATGGATATCTAAATAAAAGTAATTTTCTA


CTTACTGATAAGCCCGAGGATATAAAAGGAAATGGGGAGAGTTACTTTTTGTTTCCCTTAGATAGTTATCAA


ACACAGATAATCTCTGGAAACTTACAAAAACTTCATTATTTAGATTTAAATCTTAATTACCCTAAAGGTACA


TTTTATCGAAATGGACCAGTAAGAGAACATGGAACACCAACCAAACTTTATATAAATAGTTTAAAACAGAAA


AATTATGACATCTTTAATTTTGGTATAGATATATCTGGTTTTAGACAAGTTTATAATGAGGATTATAAGAAA


AATCAAGATGGTACTTTTCAAAAATTGAAAGAGGAAGCTTTTGAACTTTCAGATGGGGAAATAACAGAACTA


ATGAAGTCATTCTCTTCTAAACCTGAGTATTATACCCCGATAGTAACTTCATCCGATGCATCTAACAATGAA


ATTTTATCTAAAATTCAGCAACAATTTGAAAAGATTTTAACAAAAGAAAACTCAATTGTTAATGGAACTATA


GAAGATCCTATGGGTGACAAAATCAATTTACAGCTTGGCAACGGACAAACATTGCAACCAAGTGATTATACT


TTACAGGGAAATGATGGAAGTATAATGAAAGATAGCATTGCAACTGGTGGGCCTAATAATGATGGTGGAATA


CTTAAAGGGGTTAAATTAGAATACATCAAAAATAAACTCTACGTTAGAGGTTTGAACTTAGGGGAGGGACAA


AAAGTAACACTCACATATGATGTGAAACTAGATGACAGTTTTATAAGTAACAAATTCTATGACACTAATGGT


AGAACAACATTGAATCCTAAATCAGAGGATCCTAATACACTTAGAGATTTTCCAATCCCTAAAATTCGTGAT


GTGAGAGAATATCCTACAATAACGATTAAAAACGAGAAGAAGTTAGGTGAAATTGAATTTACAAAAGTTGAT


AAAGATAATAATAAGTTGCTTCTCAAAGGAGCTACGTTTGAACTTCAAGAATTTAATGAAGATTATAAACTT


TATTTACCAATAAAAAATAATAATTCAAAAGTAGTGACGGGAGAAAACGGCAAAATTTCTTACAAAGATTTG


AAAGATGGCAAATATCAGTTAATAGAAGCAGTTTCGCCGAAGGATTATCAAAAAATTACTAATAAACCAATT


TTAACTTTTGAAGTTGTTAAAGGATCGATACAAAATATAATAGCTGTTAATAAACAGATTTCTGAATATCAT


GAGGAAGGTGACAAGCATTTAATTACCAACACGCATATTCCACCAAAAGGAATTATTCCGATGACAGGTGGG


AAAGGAATTCTATCTTTCATTTTAATAGGTGGATCTATGATGTCTATTGCAGGTGGAATTTATATTTGGAAA


AGATATAAGAAATCTAGTGATATATCTAGAGAAAAAGATTAA


 


ORF amino acid sequence (SEQ ID NO: 201):


MRKYQKFSKILTLSLFCLSQIPLNTNVLGESTVPENGAKGKLVVKKTDDQNKPLSKATFVLKTTAHPESKIE


KVTAELTGEATFDNLIPGDYTLSEETAPEGYKKTNQTWQVKVESNGKTTIQNSGDKNSTIGQNQEELDKQYP


PTGIYEDTKESYKLEHVKGSVPNGKSEAKAVNPYSSEGEHIREIPEGTLSKRISEVGDLAHNKYKIELTVSG


KTIVKPVDKQKPLDVVFVLDNSNSMNNDGPNFQRHNKAKKAAEALGTAVKDILGANSDNRVALVTYGSDIFD


GRSVDVVKGFKEDDKYYGLQTKFTIQTENYSHKQLTNNAEEIIKRIPTEAPKAKWGSTTNGLTPEQQKEYYL


SKVGETFTMKAFMEADDILSQVNRNSQKIIVHVTDGVPTRSYAINNFKLGASYESQFEQMKKNGYLNKSNFL


LTDKPEDIKGNGESYFLFPLDSYQTQIISGNLQKLHYLDLNLNYPKGTFYRNGPVREHGTPTKLYINSLKQK


NYDIFNFGIDISGFRQVYNEDYKKNQDGTFQKLKEEAFELSDGEITELMKSFSSKPEYYTPIVTSSDASNNE


ILSKIQQQFEKILTKENSIVNGTIEDPMGDKINLQLGNGQTLQPSDYTLQGNDGSIMKDSIATGGPNNDGGI


LKGVKLEYIKNKLYVRGLNLGEGQKVTLTYDVKLDDSFISNKFYDTNGRTTLNPKSEDPNTLRDFPIPKIRD


VREYPTITIKNEKKLGEIEFTKVDKDNNKLLLKGATFELQEFNEDYKLYLPIKNNNSKVVTGENGKISYKDL


KDGKYQLIEAVSPKDYQKITNKPILTFEVVKGSIQNIIAVNKQISEYHEEGDKHLITNTHIPPKGIIPMTGG


KGILSFILIGGSMMSIAGGIYIWKRYKKSSDISREKD


 


Strain IC306


ORF DNA sequence (SEQ ID NO: 221):


ATGAGAAAATACCAAAAATTTTCTAAAATATTGACGTTAAGTCTTTTTTGTTTGTCGCAAATACCGCTTAAT


ACCAATGTTTTAGGGGAAAGTACCGTACCGGAAAATGGTGCTAAAGGAAAGTTAGTTGTTAAAAAGACAGAT


GACCAGAACAAACCACTTTCAAAAGCTACCTTTGTTTTAAAAACTACTGCTCATCCAGAAAGTAAAATAGAA


AAAGTAACTGCTGAGCTAACAGGTGAAGCTACTTTTGATAATCTCATACCTGGAGATTATACTTTATCAGAA


GAAACAGCGCCCGAAGGTTATAAAAAGACTAACCAGACTTGGCAAGTTAAGGTTGAGAGTAATGGAAAAACT


ACGATACAAAATAGTGGTGATAAAAATTCCACAATTGGACAAAATCAGGAAGAACTAGATAAGCAGTATCCC


CCCACAGGAATTTATGAAGATACAAAGGAATCTTATAAACTTGAGCATGTTAAAGGTTCAGTTCCAAATGGA


AAGTCAGAGGCAAAAGCAGTTAACCCATATTCAAGTGAAGGTGAGCATATAAGAGAAATTCCAGAGGGAACA


TTATCTAAACGTATTTCAGAAGTAGGTGATTTAGCTCATAATAAATATAAAATTGAGTTAACTGTCAGTGGA


AAAACCATAGTAAAACCAGTGGACAAACAAAAGCCGTTAGATGTTGTCTTCGTACTCGATAATTCTAACTCA


ATGAATAACGATGGCCCAAATTTTCAAAGGCATAATAAAGCCAAGAAAGCTGCCGAAGCTCTTGGGACCGCA


GTAAAAGATATTTTAGGAGCAAACAGTGATAATAGGGTTGCATTAGTTACCTATGGTTCAGATATTTTTGAT


GGTAGGAGTGTAGATGTCGTAAAAGGATTTAAAGAAGATGATAAATATTATGGCCTTCAAACTAAGTTCACA


ATTCAGACAGAGAATTATAGTCATAAACAATTAACAAATAATGCTGAAGAGATTATAAAAAGGATTCCGACA


GAAGCTCCTAAAGCTAAGTGGGGATCTACTACCAATGGATTAACTCCAGAGCAACAAAAGGAGTACTATCTT


AGTAAAGTAGGAGAAACATTTACTATGAAAGCCTTCATGGAGGCAGATGATATTTTGAGTCAAGTAAATCGA


AATAGTCAAAAAATTATTGTTCATGTAACTGATGGTGTTCCTACGAGATCATATGCTATTAATAATTTTAAA


CTGGGTGCATCATATGAAAGCCAATTTGAACAAATGAAAAAAAATGGATATCTAAATAAAAGTAATTTTCTA


CTTACTGATAAGCCCGAGGATATAAAAGGAAATGGGGAGAGTTACTTTTTGTTTCCCTTAGATAGTTATCAA


ACACAGATAATCTCTGGAAACTTACAAAAACTTCATTATTTAGATTTAAATCTTAATTACCCTAAAGGTACA


TTTTATCGAAATGGACCAGTAAGAGAACATGGAACACCAACCAAACTTTATATAAATAGTTTAAAACAGAAA


AATTATGACATCTTTAATTTTGGTATAGATATATCTGGTTTTAGACAAGTTTATAATGAGGATTATAAGAAA


AATCAAGATGGTACTTTTCAAAAATTGAAAGAGGAAGCTTTTGAACTTTCAGATGGGGAAATAACAGAACTA


ATGAAGTCATTCTCTTCTAAACCTGAGTATTATACCCCGATAGTAACTTCATCCGATGCATCTAACAATGAA


ATTTTATCTAAAATTCAGCAACAATTTGAAAAGATTTTAACAAAAGAAAACTCAATTGTTAATGGAACTATA


GAAGATCCTATGGGTGACAAAATCAATTTACAGCTTGGCAACGGACAAACATTGCAACCAAGTGATTATACT


TTACAGGGAAATGATGGAAGTATAATGAAAGATAGCATTGCAACTGGTGGGCCTAATAATGATGGTGGAATA


CTTAAAGGGGTTAAATTAGAATACATCAAAAATAAACTCTACGTTAGAGGTTTGAACTTAGGGGAGGGACAA


AAAGTAACACTCACATATGATGTGAAACTAGATGACAGTTTTATAAGTAACAAATTCTATGACACTAATGGT


AGAACAACATTGAATCCTAAATCAGAGGATCCTAATACACTTAGAGATTTTCCAATCCCTAAAATTCGTGAT


GTGAGAGAATATCCTACAATAACGATTAAAAACGAGAAGAAGTTAGGTGAAATTGAATTTACAAAAGTTGAT


AAAGATAATAATAAGTTGCTTCTCAAAGGAGCTACGTTTGAACTTCAAGAATTTAATGAAGATTATAAACTT


TATTTACCAATAAAAAATAATAATTCAAAAGTAGTGACGGGAGAAAACGGCAAAATTTCTTACAAAGATTTG


AAAGATGGCAAATATCAGTTAATAGAAGCAGTTTCGCCGAAGGATTATCAAAAAATTACTAATAAACCAATT


TTAACTTTTGAAGTTGTTAAAGGATCGATACAAAATATAATAGCTGTTAATAAACAGATTTCTGAATATCAT


GAGGAAGGTGACAAGCATTTAATTACCAACACGCATATTCCACCAAAAGGAATTATTCCGATGACAGGTGGG


AAAGGAATTCTATCTTTCATTTTAATAGGTGGATCTATGATGTCTATTGCAGGTGGAATTTATATTTGGAAA


AGATATAAGAAATCTAGTGATATATCTAGAGAAAAAGATTAA


 


ORF amino acid sequence (SEQ ID NO: 202):


MRKYQKFSKILTLSLFCLSQIPLNTNVLGESTVPENGAKGKLVVKKTDDQNKPLSKATFVLKTTAHPESKIE


KVTAELTGEATFDNLIPGDYTLSEETAPEGYKKTNQTWQVKVESNGKTTIQNSGDKNSTIGQNQEELDKQYP


PTGIYEDTKESYKLEHVKGSVPNGKSEAKAVNPYSSEGEHIREIPEGTLSKRISEVGDLAHNKYKIELTVSG


KTIVKPVDKQKPLDVVFVLDNSNSMNNDGPNFQRHNKAKKAAEALGTAVKDILGANSDNRVALVTYGSDIFD


GRSVDVVKGFKEDDKYYGLQTKFTIQTENYSHKQLTNNAEEIIKRIPTEAPKAKWGSTTNGLTPEQQKEYYL


SKVGETFTMKAFMEADDILSQVNRNSQKIIVHVTDGVPTRSYAINNFKLGASYESQFEQMKKNGYLNKSNFL


LTDKPEDIKGNGESYFLFPLDSYQTQIISGNLQKLHYLDLNLNYPKGTFYRNGPVREHGTPTKLYINSLKQK


NYDIFNFGIDISGFRQVYNEDYKKNQDGTFQKLKEEAFELSDGEITELMKSFSSKPEYYTPIVTSSDASNNE


ILSKIQQQFEKILTKENSIVNGTIEDPMGDKINLQLGNGQTLQPSDYTLQGNDGSIMKDSIATGGPNNDGGI


LKGVKLEYIKNKLYVRGLNLGEGQKVTLTYDVKLDDSFISNKFYDTNGRTTLNPKSEDPNTLRDFPIPKIRD


VREYPTITIKNEKKLGEIEFTKVDKDNNKLLLKGATFELQEFNEDYKLYLPIKNNNSKVVTGENGKISYKDL


KDGKYQLIEAVSPKDYQKITNKPILTFEVVKGSIQNIIAVNKQISEYHEEGDKHLITNTHIPPKGIIPMTGG


KGILSFILIGGSMMSIAGGIYIWKRYKKSSDISREKD


 


Strain IC458


ORF DNA sequence (SEQ ID NO: 222):


ATGAGAAAATACCAAAAATTTTCTAAAATATTGACGTTAAGTCTTTTTTGTTTGTCGCAAATACCGCTTAAT


ACCAATGTTTTAGGGGAAAGTACCGTACCGGAAAATGGTGCTAAAGGAAAGTTAGTTGTTAAAAAGACAGAT


GACCAGAACAAACCACTTTCAAAAGCTACCTTTGTTTTAAAAACTACTGCTCATCCAGAAAGTAAAATAGAA


AAAGTAACTGCTGAGGTAACAGGTGAAGCTACTTTTGATAATCTCACACCTGGAGATTACACTTTATCAGAA


GAAACGGCACCCGAAGGATACAAAAAGACTACCCAGACTTGGCAAGTTAAGGTTGAGAGTAATGGAAAAACT


ACGATACAAAATAGTGATGATAAAAAATCTATAATTGAACAAAGGCAAGAGGAACTAGATAAGCAGTATCCC


CTTACAGGAGCTTATGAAGATACAAAAGAATCTTATAATCTTGAGCATGTTAAAAATTCAATTCCAAATGGG


AAATTAGAGGCAAAAGCAGTTAATCCATATTCAAGTGAAGGTGAGCACATAAGAGAAATTCAAGAGGGAACA


TTATCTAAACGTATTTCAGAAGTAAATGATTTGGATCATAATAAATATAAAATTGAGTTAACTGTTAGCGGT


AAATCCATAATAAAAACTATAAATAAAGATGAACCTCTGGATGTTGTTTTTGTTCTTGATAATTCAAATTCT


ATGAAGAATAATGGAAAAAATAACAAGGCAAAAAAGGCAGGTGAAGCAGTAGAAACAATTATAAAAGATGTT


TTAGGAGCAAATGTTGAAAACCGAGCAGCTTTAGTTACTTATGGTTCAGATATTTTTGATGGAAGGACAGTT


AAAGTTATAAAAGGTTTTAAAGAGGATCCTTATCATGGACTTGAAACTAGTTTCACAGTTCAGACAAATGAT


TATAGCTATAAAAAGTTCACTAATATTGCTGCTGATATTATAAAAAAGATCCCTAAAGAAGCTCCAGAAGCT


AAGTGGGGGGGGACAAGTCTAGGATTAACTCCAGAAAAAAAGAGGGAATATGATTTAAGTAAAGTAGGTGAG


ACCTTTACAATGAAAGCTTTTATGGAGGCAGATACCTTGTTAAGTAGTATACAGCGTAAGAGTAGAAAGATT


ATTGTTCATCTAACTGACGGTGTTCCAACAAGATCATATGCTATTAATAGTTTTGTAACAGGTTCAACATAC


GCAAATCAATTTGAGAGAATAAAAGAAAAAGGTTATTTAGACAAAAATAATTATTTTATAACTGATGATCCA


GAAAAGATCAAAGGCAATGGGGAGAGTTACTTTTTGTTTCCCTTAGATAGTTATCAAACACAGATAATTTCT


GGAAACTTACAAAAACTTCATTATTTAGATTTAAATCTTAATTACCCTAAAGGTACAATTTATAGAAATGGA


CCAGTAAGAGAACATGGAACACCAACCAAACTTTATATAAATAGTTTAAAACAGAAAAATTATGACATCTTT


AATTTTGGTATAGATATATCTGGTTTTAGACAAGTTTATAATGAGGATTATAAGAAAAATCAAGATGGTACT


TTTCAAAAATTGAAAGAGGAAGCTTTTGAACTTTCAGGTGGGGAAATAACAGAACTAATGAAGTCATTCTCT


TCTAAACCTGAGTATTATACCCCGATAGTAACTTCAGCTGATGTATCTAATAATGAAATTTTATCTAAAATT


CAGCAACAATTTGAAAAGATTTTAACAAAGGAAAACTCAATTGTTAATGGAACTATAGAAGATCCTATGGGT


GATAAAATCAATTTACAGCTTGGCAACGGACAAACATTGCAACCAAGTGATTATACTTTACAGGGAAATGAT


GGAAGTATAATGAAAGATAGCATTGCAACTGGTGGGCCTAATAATGATGGCGGGATACTTAAAGGGGTTAAA


TTAGAATACATCAAAAATAAACTCTACGTTAGAGGTTTGAACTTAGGGGAGGGGCAAAAAGTAACACTCACA


TATGATGTGAAACTAGATGACAGTTTTATTAGTAACAAATTCTATGACACTAATGGTAGAACAACATTGAAT


CCTAAATCAGAGGAACCTGATACACTTAGAGATTTTCCAATCCCTAAAATTCGTGATGTGAGAGAATATCCT


ACAATAACGATTAAAAACGAGAAGAAGTTAGGTGAAATTGAATTTACAAAAGTTGATAAAGATAATAATAAG


TTGCTTCTCAAAGGAGCTACATTTGAACTTCAAGAATTTAATGAAGATTATAAACTTTATTTACCAATAAAA


AATAATAATTCAAAAGTAGTGACGGGAGAAAACGGCAAAATTTCTTACAAAGATTTGAAAGATGGCAAATAT


CAGTTAATAGAAGCAGTTTCGCCGAAGGATTATCAAAAAATTACTAATAAACCAATTTTAACTTTTGAAGTT


GTTAAAGGATCGATACAAAATATAATAGCTGTTAATAAACAGATTTCTGAATATCATGAGGAAGGTGACAAG


CATTTAATTACCAACACGCATATTCCACCAAAAGGAATTATTCCGATGACAGGTGGGAAAGGAATTCTATCT


TTCATTTTAATAGGTGGAGCTATGATGTCTATTGCAGGTGGAATTTATATTTGGAAAAAACATAAGAAATCT


AGTGATGCATCAATCGAGAAAGATTAA


 


ORF amino acid sequence (SEQ ID NO: 203):


MRKYQKFSKILTLSLFCLSQIPLNTNVLGESTVPENGAKGKLVVKKTDDQNKPLSKATFVLKTTAHPESKIE


KVTAEVTGEATFDNLTPGDYTLSEETAPEGYKKTTQTWQVKVESNGKTTIQNSDDKKSIIEQRQEELDKQYP


LTGAYEDTKESYNLEHVKNSIPNGKLEAKAVNPYSSEGEHIREIQEGTLSKRISEVNDLDHNKYKIELTVSG


KSIIKTINKDEPLDVVFVLDNSNSMKNNGKNNKAKKAGEAVETIIKDVLGANVENRAALVTYGSDIFDGRTV


KVIKGFKEDPYHGLETSFTVQTNDYSYKKFTNIAADIIKKIPKEAPEAKWGGTSLGLTPEKKREYDLSKVGE


TFTMKAFMEADTLLSSIQRKSRKIIVHLTDGVPTRSYAINSFVTGSTYANQFERIKEKGYLDKNNYFITDDP


EKIKGNGESYFLFPLDSYQTQIISGNLQKLHYLDLNLNYPKGTIYRNGPVREHGTPTKLYINSLKQKNYDIF


NFGIDISGFRQVYNEDYKKNQDGTFQKLKEEAFELSGGEITELMKSFSSKPEYYTPIVTSADVSNNEILSKI


QQQFEKILTKENSIVNGTIEDPMGDKINLQLGNGQTLQPSDYTLQGNDGSIMKDSIATGGPNNDGGILKGVK


LEYIKNKLYVRGLNLGEGQKVTLTYDVKLDDSFISNKFYDTNGRTTLNPKSEEPDTLRDFPIPKIRDVREYP


TITIKNEKKLGEIEFTKVDKDNNKLLLKGATFELQEFNEDYKLYLPIKNNNSKVVTGENGKISYKDLKDGKY


QLIEAVSPKDYQKITNKPILTFEVVKGSIQNIIAVNKQISEYHEEGDKHLITNTHIPPKGIIPMTGGKGILS


FILIGGAMMSIAGGIYIWKKHKKSSDASIEKD
















TABLE 12





Amino acid and encoding DNA sequences of gbs2018 proteins derived from


different strains of S. agalactiae







Strain 0176H4A


ORF DNA sequence (SEQ ID NO: 180):


ATGAATAATAACGAAAAAAAAGTAAAATACTTTTTAAGAAAAACAGCTTATGGTTTGGCCTCAATGTCAGCA


GCGTTTGCTGTATGTAGTGGTATTGTACACGCGGATACTAGTTCAGGAATATCGGATTCAATTCCTCATAAG


AAACAAGTTAATTTAGGGGCGGTTACTCTGAAGAATTTGATTTCTAAATATCGTGGTAATGACAAAGCTATT


GCTATACTTCTAAGTAGAGTAGATGATTTTAATAGAGCATCACAGGATACACTTCCACAATTAATTAATAGT


ACTGAAGCAGAAATTAACAATACTTTACCTCAGGGACGAATTATTAAACAGAGTATACCAGTCGTAAGATTA


AAAGTTGAGAGATTGGGAAGTGGTGCAATTAAGGCTGAGTCGATTAATAATATTAAAGCTGAATCAATTAAT


AAAATTCAGGGTAAATCAACTAATACAATTAAGGCTGAGTCCATTAATAAAATTAAAGTAGAGTCTATTAAT


ACAATCAAAGCCGAATCAATTAATAAAATTCAAGCTAAGCCAATTAACACAATCAAAGCCGAGTCTATTAAT


ACAATTAAGGCTGAATCAATTCATAAAATTAAACCTCAATCAATAAAAAGTACTAGTGCTACACATGTTAAA


GTTAGTGATCAAGAACTAGCTAAGCAGTCAAGACGTTCTCAAGATATCATTAAATCATTAGGTTTCCTTTCA


TCAGACCAAAAAGATATTTTAGTTAAATCTATTAGCTCTTCAAAAGATTCGCAACTTATTCTTAAATTTGTA


ACACAAGCCACGCAACTGAATAATGCTGAATCAACAAAAGCTAAGCACATGGCTCAAAATGACGTGGCTTCA


ATAAAAAATATAAGCCTCGAAGTCTTAGAAGAATATAAAGAAAAAATTCAAAGAGCTAGCACTAAGAGTCAA


GTTGATGAGCTTGTAGCAGAAGCTAAAAAAGTTGTTAATTCCAATAAAGAAACATTGGTAAATCAGGCCAAT


GGTAAAAAGCAAGAAATTGCTAAGTTAGAAAATTTATCTAACGATGAAATGTTGAGATATAATACTGCAATT


GATAATGTAGTGAAACAGTATAATGAAGGTAAGCTCAATATTACTGATGCAATGAATGCTTTAAATAGTATT


AAGCAAGCAGCACAGGAAGTTGCCCAGAAAAACTTACAAAAGCAGTATGCTAAAAAAATTGAAAGAATAAGT


TTAAAAGGATTAGCGTTATCCAAAAAGGCTAAAGAAATTTATGAAAAGCATAAAAGTATTTTGCCTACACCT


GGATATTATGCAGACTCTGTGGGAACTTATTTGAATAGGTTTAGAGATAAACGAACTTTCGGAAATAGAAGT


GTTTGGACTGGTCAAAGTGGACTTGATGAAGCAAAAAAAATGCTTGATGAAGTCAAAAAGCTTTTAAAAGAA


CTTCAAGACCTTACCAGAGGTACTAAAGAAGATAAAAAACCAGACGTTAAGCCAGAAGCCAAACCAGAGGCC


AAACCAAATATTCAAGTACCTAAACAAGCACCTACAGAAGCTGCAAAACCAGCTTTGTCACCAGAAGCCTTG


ACAAGATTGACTACATGGTATAATCAAGCTAAAGATCTGCTTAAAGATGATCAAGTAAAGGACAAATATGTA


GATATACTTGCAGTTCAAAAAGCTGTTGACCAAGCTTATGATCATGTGGAAGAGGGAAAATTTATTACCACT


GATCAAGCAAATCAATTAGCTAACAAGCTACGTGATGCTTTACAAAGTTTAGAATTAAAAGATAAAAAAGTA


GCCAAACCAGAAGCTAAGCCAGAAGTTAAACCAGAAGCCAAACCAGATGTTAAGCCAGACGTTAAGCCAGAA


GCTAAGCCAGAAGCCAAACCAGAGGCCAAACCAGAAGCCAAACCAGAGGCCAAACCAGAAGCCAAACCAGAG


GCTAAGCCAGAAGTTAAACCAGACGTTAAGCCAGAGGCCAAACCAGACGTTAAGCCAGAAGCCAAACCAGAC


GTTAAGCCAGAGGCTAAGCCAGAAGTTAAACCAGACGTTAAGCCAGAAGTTAAACCAGAGGCTAAACCAGAA


ATTAAACCAGACGTTAAGCCAGAGGCCAGACCAGAGGCTAAGCCAGAAGTTAAACCAGACGTTAAGCCAGAG


GCCAAACCAGAAGTTAAACCAGACGTTAAGCCAGAGGCCAAACCAGAGGCTAAGCCAGAAGTTAAACCAGAC


GTTAAGCCAGAGGCTAAACCAGAAGCCAAACCAGCAACCAAAAAATCGGTTAATACTAGCGGAAACTTGGCG


GCTAAAAAAGCTATTGAAAACAAAAAGTATAGTAAAAAATTACCATCAACGGGTGAAGCCGCAAGTCCACTC


TTAGCAATTGTATCACTAATTGTTATGTTAAGTGCAGGTCTTATTACGATAGTTTTAAAGCATAAAAAAAAT


TAA


 


ORF amino acid sequence (SEQ ID NO: 175):


MNNNEKKVKYFLRKTAYGLASMSAAFAVCSGIVHADTSSGISDSIPHKKQVNLGAVTLKNLISKYRGNDKAI


AILLSRVDDFNRASQDTLPQLINSTEAEINNTLPQGRIIKQSIPVVRLKVERLGSGAIKAESINNIKAESIN


KIQGKSTNTIKAESINKIKVESINTIKAESINKIQAKPINTIKAESINTIKAESIHKIKPQSIKSTSATHVK


VSDQELAKQSRRSQDIIKSLGFLSSDQKDILVKSISSSKDSQLILKFVTQATQLNNAESTKAKHMAQNDVAS


IKNISLEVLEEYKEKIQRASTKSQVDELVAEAKKVVNSNKETLVNQANGKKQEIAKLENLSNDEMLRYNTAI


DNVVKQYNEGKLNITDAMNALNSIKQAAQEVAQKNLQKQYAKKIERISLKGLALSKKAKEIYEKHKSILPTP


GYYADSVGTYLNRFRDKRTFGNRSVWTGQSGLDEAKKMLDEVKKLLKELQDLTRGTKEDKKPDVKPEAKPEA


KPNIQVPKQAPTEAAKPALSPEALTRLTTWYNQAKDLLKDDQVKDKYVDILAVQKAVDQAYDHVEEGKFITT


DQANQLANKLRDALQSLELKDKKVAKPEAKPEVKPEAKPDVKPDVKPEAKPEAKPEAKPEAKPEAKPEAKPE


AKPEVKPDVKPEAKPDVKPEAKPDVKPEAKPEVKPDVKPEVKPEAKPEIKPDVKPEARPEAKPEVKPDVKPE


AKPEVKPDVKPEAKPEAKPEVKPDVKPEAKPEAKPATKKSVNTSGNLAAKKAIENKKYSKKLPSTGEAASPL


LAIVSLIVMLSAGLITIVLKHKKN


 


Strain 12401


ORF DNA sequence (SEQ ID NO: 181):


ATGAATAATAACGAAAAAAAAGTAAAATACTTTTTAAGAAAAACAGCTTATGGTTTGGCCTCAATGTCAGCA


GCGTTTGCTGTATGTAGTGGTATTGTACACGCGGATACTAGTTCAGGAATATCGGATTCAATTCCTCATAAG


AAACAAGTTAATTTAGGGGCGGTTACTCTGAAGAATTTGATTTCTAAATATCGTGGTAATGACAAAGCTATT


GCTATACTTTTAAGTAGAGTAAATGATTTTAATAGAGCATCACAGGATACACTTCCACAATTAATTAATAGT


ACTGAAGCAGAAATTAACAATACTTTACCTCAGGGACGAATTATTAAACAGAGTATACCAGTCGTAAGATTA


AAAGTTGAGAGATTGGGAAGTGGTGCAATTAAGGCTGAGTCGATTAATAATATTAAAGCTGAATCAATTAAT


AAAATTCAGGGTAAATCAACTAATACAATTAAGGCTGAGTCCATTAATAAAATTAAAGTAGAGTCTATTAAT


ACAATCAAAGCCGAATCAATTAATAAAATTCAAGCTAAGCCAATTAACACAATCAAAGCCGAGTCTATTAAT


ACAATTAAGGCTGAATCAATTCATAAAATTAAACCTCAATCAATAAAAAGTACTAGTGCTACACATGTTAAA


GTTAGTGATCAAGAACTAGGTAAGCAGTCAAGACGTTCTCAAGATATCATTAAATCATTAGGTTTCCTTTCA


TCAGACCAAAAAGATATTTTAGTTAAATCTATTAGCTCTTCAAAAGATTCGCAACTTATTCTTAAATTTGTA


ACACAAGCCACGCAACTGAATAATGCTGAATCAACAAAAGCTAAGCACATGGCTCAAAATGACGTGGCTTCA


ATAAAAAATATAAGCCTCGAAGTCTTAGAAGAATATAAAGAAAAAATTCAAAGAGCTAGCACTAAGAGTCAA


GTTGATGAGCTTGTAGCAGAAGCTAAAAAAGTTGTTAATTCCAATAAAGAAACATTGGTAAATCAGGCCAAT


GGTAAAAAGCAAGAAATTGCTAAGTTAGAAAATTTATCTAACGATGAAATGTTGAGATATAATACTGCAATT


GATAATGTAGTGAAACAGTATAATGAAGGTAAGCTCAATATTACTGATGCAATGAATGCTTTAAATAGTATT


AAGCAAGCAGCACAGGAAGTTGCCCAGAAAAACTTACAAAAGCAGTATGCTAAAAAAATTGAAAGAATAAGT


TTAAAAGGATTAGCGTTATCCAAAAAGGCTAAAGAAATTTATGAAAAGCATAAAAGTATTTTGCCTACACCT


GGATATTATGCAGACTCTGTGGGAACTTATTTGAATAGGTTTAGAGATAAACGAACTTTCGGAAATAGAAGT


GTTTGGACTGGTCAAAGTGGACTTGATGAAGCAAAAAAAATGCTTGATGAAGTCAAAAAGCTTTTAAAAGAA


CTTCAAGACCTTACCAGAGGTACTAAAGAAGATAAAAAACCAGACGTTAAGCCAGAAGCCAAACCAGAGGCC


AAACCAAATATTCAAGTACCTAAACAAGCACCTACAGAAGCTGCAAAACCAGCTTTGTCACCAGAAGCCTTG


ACAAGATTGACTACATGGTATAATCAAGCTAAAGATCTGCTTAAAGATGATCAAGTAAAGGACAAATATGTA


GATATACTTGCAGTTCAAAAAGCTGTTGACCAAGCTTATGATCATGTGGAAGAGGGAAAATTTATTACCACT


GATCAAGCAAATCAATTAGCTAACAAGCTACGTGATGCTTTACAAAGTTTAGAATTAAAAGATAAAAAAGTA


GCCAAACCAGAAGCTAAGCCAGAAGTTAAACCAGAAGCCAAACCAGAGGCCAAACCAGAAGCCAAACCAGAG


GCTAAGCCAGAAGTTAAACCAGACGTTAAGCCAGAGGCCAAACCAGACGTTAAGCCAGAAGCCAAACCAGAC


GTTAAGCCAGAAGTTAAACCAGACGTTAAGCCAGAGGCCAAACCAGACGTTAAGCCAGAAGCTAAGCCAGAC


GTTAAGCCAGAAGTTAAACCAGAGGCTAAGCCAGAAGCCAAACCAGAGGCTAAACCAGAAATTAAACCAGAC


GTTAAGCCAGAGGCCAGACCAGAGGCTAAGCCAGAAGTTAAACCAGACGTTAAGCCAGAGGCCAAACCAGAG


GTTAAGCCAGAAGTTAAACCAGACGTTAAGCCAGAGGCTAAACCAGAAGCCAAACCAGCAACCAAAAAATCG


GTTAATACTAGCGGAAACTTGGCGGTTAAAAAAGCTATTGAAAACAAAAAGTATAGTAAAAAATTACCATCA


ACGGGTGAAGCCGCAAGTCCACTCTTAGCAATTGTATCACTAATTGTTATGTTAAGTGCAGGTCTTATTACG


ATAGTTTTAAAGCATAAAAAAAATTAA


 


ORF amino acid sequence (SEQ ID NO: 176):


MNNNEKKVKYFLRKTAYGLASMSAAFAVCSGIVHADTSSGISDSIPHKKQVNLGAVTLKNLISKYRGNDKAI


AILLSRVNDFNRASQDTLPQLINSTEAEINNTLPQGRIIKQSIPVVRLKVERLGSGAIKAESINNIKAESIN


KIQGKSTNTIKAESINKIKVESINTIKAESINKIQAKPINTIKAESINTIKAESIHKIKPQSIKSTSATHVK


VSDQELGKQSRRSQDIIKSLGFLSSDQKDILVKSISSSKDSQLILKFVTQATQLNNAESTKAKHMAQNDVAS


IKNISLEVLEEYKEKIQRASTKSQVDELVAEAKKVVNSNKETLVNQANGKKQEIAKLENLSNDEMLRYNTAI


DNVVKQYNEGKLNITDAMNALNSIKQAAQEVAQKNLQKQYAKKIERISLKGLALSKKAKEIYEKHKSILPTP


GYYADSVGTYLNRFRDKRTFGNRSVWTGQSGLDEAKKMLDEVKKLLKELQDLTRGTKEDKKPDVKPEAKPEA


KPNIQVPKQAPTEAAKPALSPEALTRLTTWYNQAKDLLKDDQVKDKYVDILAVQKAVDQAYDHVEEGKFITT


DQANQLANKLRDALQSLELKDKKVAKPEAKPEVKPEAKPEAKPEAKPEAKPEVKPDVKPEAKPDVKPEAKPD


VKPEVKPDVKPEAKPDVKPEAKPDVKPEVKPEAKPEAKPEAKPEIKPDVKPEARPEAKPEVKPDVKPEAKPE


VKPEVKPDVKPEAKPEAKPATKKSVNTSGNLAVKKAIENKKYSKKLPSTGEAASPLLAIVSLIVMLSAGLIT


IVLKHKKN


 


Strain BAA23


ORF DNA sequence (SEQ ID NO: 182):


ATGAATAATAACGAAAAAAAAGTAAAATACTTTTTAAGAAAAACAGCTTATGGTTTGGCCTCAATGTCAGCA


GCGTTTGCTGTATGTAGTGGTATTGTACACGCGGATACTAGTTCAGGAATATCGGATTCAATTCCTCATAAG


AAACAAGTTAATTTAGGGGCGGTTACTCTGAAGAATTTGATTTCTAAATATCGTGGTAATGACAAAGCTATT


GCTATACTTCTAAGTAGAGTAGATGATTTTAATAGAGCATCACAGGATACACTTCCACAATTAATTAATAGT


ACTGAAGCAGAAATTAACAATACTTTACCTCAGGGACGAATTATTAAACAGAGTATACCAGTCGTAAGATTA


AAAGTTGAGAGATTGGGAAGTGGTGCAATTAAGGCTGAGTCGATTAATAATATTAAAGCTGAATCAATTAAT


AAAATTCAGGGTAAATCAACTAATACAATTAAGGCTGAGTCCATTAATAAAATTAAAGTAGAGTCTATTAAT


ACAATCAAAGCCGAATCAATTAATAAAATTCAAGCTAAGCCAATTAACACAATCAAAGCCGAGTCTATTAAT


ACAATTAAGGCTGAATCAATTCATAAAATTAAACCTCAATCAATAAAAAGTACTAGTGCTACACATGTTAAA


GTTAGTGATCAAGAACTAGCTAAGCAGTCAAGACGTTCTCAAGATATCATTAAATCATTAGGTTTCCTTTCA


TCAGACCAAAAAGATATTTTAGTTAAATCTATTAGCTCTTCAAAAGATTCGCAACTTATTCTTAAATTTGTA


ACACAAGCCACGCAACTGAATAATGCTGAATCAACAAAAGCTAAGCACATGGCTCAAAATGACGTGGCTTCA


ATAAAAAATATAAGCCTCGAAGTCTTAGAAGAATATAAAGAAAAAATTCAAAGAGCTAGCACTAAGAGTCAA


GTTGATGAGCTTGTAGCAGAAGCTAAAAAAGTTGTTAATTCCAATAAAGAAACATTGGTAAATCAGGCCAAT


GGTAAAAAGCAAGAAATTGCTAAGTTAGAAAATTTATCTAACGATGAAATGTTGAGATATAATACTGCAATT


GATAATGTAGTGAAACAGTATAATGAAGGTAAGCTCAATATTACTGATGCAATGAATGCTTTAAATAGTATT


AAGCAAGCAGCACAGGAAGTTGCCCAGAAAAACTTACAAAAGCAGTATGCTAAAAAAATTGAAAGAATAAGT


TTAAAAGGATTAGCGTTATCCAAAAAGGCTAAAGAAATTTATGAAAAGCATAAAAGTATTTTGCCTACACCT


GGATATTATGCAGACTCTGTGGGAACTTATTTGAATAGGTTTAGAGATAAACGAACTTTCGGAAATAGAAGT


GTTTGGACTGGTCAAAGTGGACTTGATGAAGCAAAAAAAATGCTTGATGAAGTCAAAAAGCTTTTAAAAGAA


CTTCAAGACCTTACCAGAGGTACTAAAGAAGATAAAAAACCAGACGTTAAGCCAGAAGCCAAACCAGAGGCC


AAACCAAATATTCAAGTACCTAAACAAGCACCTACAGAAGCTGCAAAACCAGCTTTGTCACCAGAAGCCTTG


ACAAGATTGACTACATGGTATAATCAAGCTAAAGATCTGCTTAAAGATGATCAAGTAAAGGACAAATATGTA


GATATATTTGCAGTTCAAAAAGCTGTTGACCAAGCTTATGATCATGTGGAAGAGGGAAAATTTATTACCACT


GATCAAGCAAATCAATTAGCTAACAAGATACGTGATGCTTTACAAAGTTTAGAATTAAAAGATAAAAAAGTA


GCCAAACCAGAGGGTAAGCCAGAAGTTAAACCAGACGTTAAGCCAGAGGCCAAACCAGACGTTAAGCCAGAA


GCCAAACCAGACGTTAAGCCAGAAGTTAAACCAGACGTTAAGCCAGAGGCCAAACCAGACGTTAAGCCAGAA


GCTAAGCCAGACGTTAAGCCAGAAGTTAAACCAGAGGCTAAGCCAGAAGTTAAACCAGACGTTAAGCCAGAG


GCCAGACCAGAGGCTAAGCCAGAAGTTAAACCAGACGTTAAGCCAGAGGCCAAACCAGAGGCTAAGCCAGAA


GTTAAACCAGACGTTAAGCCAGAGGCTAAACCAGAAGCCAAACCAGCAACCAAAAAATCGGTTAATACTAGC


GGAAACTTGGCGGTTAAAAAAGCTATTGAAAACAAAAAGTATAGTAAAAAATTACCATCAACGGGTGAAGCC


GCAAGTCCACTCTTAGCAATTGTATCACTAATTGTTATGTTAAGTGCAGGTCTTATTACGATAGTTTTAAAG


CATAAAAAAAATTAA


 


ORF amino acid sequence (SEQ ID NO: 177):


MNNNEKKVKYFLRKTAYGLASMSAAFAVCSGIVHADTSSGISDSIPHKKQVNLGAVTLKNLISKYRGNDKAI


AILLSRVDDFNRASQDTLPQLINSTEAEINNTLPQGRIIKQSIPVVRLKVERLGSGAIKAESINNIKAESIN


KIQGKSTNTIKAESINKIKVESINTIKAESINKIQAKPINTIKAESINTIKAESIHKIKPQSIKSTSATHVK


VSDQELAKQSRRSQDIIKSLGFLSSDQKDILVKSISSSKDSQLILKFVTQATQLNNAESTKAKHMAQNDVAS


IKNISLEVLEEYKEKIQRASTKSQVDELVAEAKKVVNSNKETLVNQANGKKQEIAKLENLSNDEMLRYNTAI


DNVVKQYNEGKLNITDAMNALNSIKQAAQEVAQKNLQKQYAKKIERISLKGLALSKKAKEIYEKHKSILPTP


GYYADSVGTYLNRFRDKRTFGNRSVWTGQSGLDEAKKMLDEVKKLLKELQDLTRGTKEDKKPDVKPEAKPEA


KPNIQVPKQAPTEAAKPALSPEALTRLTTWYNQAKDLLKDDQVKDKYVDIFAVQKAVDQAYDHVEEGKFITT


DQANQLANKIRDALQSLELKDKKVAKPEGKPEVKPDVKPEAKPDVKPEAKPDVKPEVKPDVKPEAKPDVKPE


AKPDVKPEVKPEAKPEVKPDVKPEARPEAKPEVKPDVKPEAKPEAKPEVKPDVKPEAKPEAKPATKKSVNTS


GNLAVKKAIENKKYSKKLPSTGEAASPLLAIVSLIVMLSAGLITIVLKHKKN


 


Strain IC105


ORF DNA sequence (SEQ ID NO: 183):


ATGAATAATAACGAAAAAAAAGTAAAATACTTTTTAAGAAAAACAGCTTATGGTTTGGCCTCAATGTCAGCA


GCGTTTGCTGTATGTAGTGGTATTGTACACGCGGATACTAGTTCAGGAATATCGGATTCAATTCCTCATAAG


AAACAAGTTAATTTAGGGGCGGTTACTCTGAAGAATTTGATTTCTAAATATCGTGGTAATGACAAAGCTATT


GCTATACTTCTAAGTAGAGTAGATGATTTTAATAGAGCATCACAGGATACACTTCCACAATTAATTAATAGT


ACTGAAGCAGAAATTAACAATACTTTACCTCAGGGACGAATTATTAAACAGAGTATACCAGTCGTAAGATTA


AAAGTTGAGAGATTGGGAAGTGGTGCAATTAAGGCTGAGTCGATTAATAATATTAAAGCTGAATCAATTAAT


AAAATTCAGGGTAAATCAACTAATACAATTAAGGCTGAGTCCATTAATAAAATTAAAGTAGAGTCTATTAAT


ACAATCAAAGCCGAATCAATTAATAAAATTCAAGCTAAGCCAATTAACACAATCAAAGCCGAGTCTATTAAT


ACAATTAAGGCTGAATCAATTCATAAAATTAAACCTCAATCAATAAAAAGTACTAGTGCTACACATGTTAAA


GTTAGTGATCAAGAACTAGGTAAGCAGTCAAGACGTTCTCAAGATATCATTAAATCATTAGGTTTCCTTTCA


TCAGACCAAAAAGATATTTTAGTTAAATCTATTAGCTCTTCAAAAGATTCGCAACTTATTCTTAAATTTGTA


ACACAAGCCACGCAACTGAATAATGCTGAATCAACAAAAGCTAAGCACATGGCTCAAAATGACGTGGCTTCA


ATAAAAAATATAAGCCTCGAAGTCTTAGAAGAATATAAAGAAAAAATTCAAAGAGCTAGCACTAAGAGTCAA


GTTGATGAGCTTGTAGCAGAAGCTAAAAAAGTTGTTAATTCCAATAAAGAAACATTGGTAAATCAGGCCAAT


GGTAAAAAGCAAGAAATTGCTAAGTTAGAAAATTTATCTAACGATGAAATGTTGAGATATAATACTGCAATT


GATAATGTAGTGAAACAGTATAATGAAGGTAAGCTCAATATTACTGATGCAATGAATGCTTTAAATAGTATT


AAGCAAGCAGCACAGGAAGTTGCCCAGAAAAACTTACAAAAGCAGTATGCTAAAAAAATTGAAAGAATAAGT


TTAAAAGGATTAGCGTTATCCAAAAAGGCTAAAGAAATTTATGAAAAGCATAAAAGTATTTTGCCTACACCT


GGATATTATGCAGACTCTGTGGGAACTTATTTGAATAGGTTTAGAGATAAACGAACTTTCGGAAATAGAAGT


GTTTGGACTGGTCAAAGTGGACTTGATGAAGCAAAAAAAATGCTTGATGAAGTCAAAAAGCTTTTAAAAGAA


CTTCAAGACCTTACCAGAGGTACTAAAGAAGATAAAAAACCAGACGTTAAGCCAGAAGCCAAACCAGAGGCC


AAACCAAATATTCAAGTACCTAAACAAGCACCTACAGAAGCTGCAAAACCAGCTTTGTCACCAGAAGCCTTG


ACAAGATTGACTACATGGTATAATCAAGCTAAAGATCTGCTTAAAGATGATCAAGTAAAGGACAAATATGTA


GATATACTTGCAGTTCAAAAAGCTGTTGACCAAGCTTATGATCATGTGGAAGAGGGAAAATTTATTACCACT


GATCAAGCAAATCAATTAGCTAACAAGCTACGTGATGCTTTACAAAGTTTAGAATTAAAAGATAAAAAAGTA


GCCAAACCAGAAGCTAAGCCAGAAGTTAAACCAGAAGCCAAACCAGATGTTAAGCCAGACGTTAAGCCAGAA


GCTAAGCCAGAAGCCAAACCAGAGGCCAAACCAGAAGCCAAACCAGAGGCCAAACCAGAAGCCAAACCAGAG


GCTAAGCCAGAAGTTAAACCAGACGTTAAGCCAGAGGCCAAACCAGACGTTAAGCCAGAAGCCAAACCAGAC


GTTAAGCCAGAGGCTAAGCCAGAAGTTAAACCAGACGTTAAGCCAGAGGCCAAACCAGACGTTAAGCCAGAA


GCTAAGCCAGACGTTAAGCCAGAAGTTAAACCAGAGGCTAAGCCAGAAGTTAAACCAGACGTTAAGCCAGAG


GCCAGACCAGAGGCTAAGCCAGAAGTTAAACCAGACGTTAAGCCAGAGGCCAAACCAGAGGCTAAGCCAGAA


GTTAAACCAGACGTTAAGCCAGAGGCTAAACCAGAAGCTAAACCAGCAACCAAAAAATCGGTTAATACTAGC


GGAAACTTGGCGGTTAAAAAAGCTATTGAAAACAAAAAGTATAGTAAAAAATTACCATCAACGGGTGAAGCC


GCAAGTCCACTCTTAGCAATTGTATCACTAATTGTTATGTTAAGTGCAGGTCTTATTACGATAGTTTTAAAG


CATAAAAAAAATTAA


 


ORF amino acid sequence (SEQ ID NO: 178):


MNNNEKKVKYFLRKTAYGLASMSAAFAVCSGIVHADTSSGISDSIPHKKQVNLGAVTLKNLISKYRGNDKAI


AILLSRVDDFNRASQDTLPQLINSTEAEINNTLPQGRIIKQSIPVVRLKVERLGSGAIKAESINNIKAESIN


KIQGKSTNTIKAESINKIKVESINTIKAESINKIQAKPINTIKAESINTIKAESIHKIKPQSIKSTSATHVK


VSDQELGKQSRRSQDIIKSLGFLSSDQKDILVKSISSSKDSQLILKFVTQATQLNNAESTKAKHMAQNDVAS


IKNISLEVLEEYKEKIQRASTKSQVDELVAEAKKVVNSNKETLVNQANGKKQEIAKLENLSNDEMLRYNTAI


DNVVKQYNEGKLNITDAMNALNSIKQAAQEVAQKNLQKQYAKKIERISLKGLALSKKAKEIYEKHKSILPTP


GYYADSVGTYLNRFRDKRTFGNRSVWTGQSGLDEAKKMLDEVKKLLKELQDLTRGTKEDKKPDVKPEAKPEA


KPNIQVPKQAPTEAAKPALSPEALTRLTTWYNQAKDLLKDDQVKDKYVDILAVQKAVDQAYDHVEEGKFITT


DQANQLANKLRDALQSLELKDKKVAKPEAKPEVKPEAKPDVKPDVKPEAKPEAKPEAKPEAKPEAKPEAKPE


AKPEVKPDVKPEAKPDVKPEAKPDVKPEAKPEVKPDVKPEAKPDVKPEAKPDVKPEVKPEAKPEVKPDVKPE


ARPEAKPEVKPDVKPEAKPEAKPEVKPDVKPEAKPEAKPATKKSVNTSGNLAVKKAIENKKYSKKLPSTGEA


ASPLLAIVSLIVMLSAGLITIVLKHKKN


 


Strain IC458


ORF DNA sequence (SEQ ID NO: 184):


ATGAATAATAACGAAAAAAAAGTAAAATACTTTTTAAGAAAAACAGCTTATGGTTTGGCCTCAATGTCAGCA


GCGTTTGCTGTATGTAGTGGTATTGTACACGCGGATACTAGTTCAGGAATATCGGCTTCAATTCCTCATAAG


AAACAAGTTAATTTAGGGGCGGTTACTCTGAAGAATTTGATTTCTAAATATCGTGGTAATGACAAAGCTATT


GCTATACTTTTAAGTAGAGTAAATGATTTTAATAGAGCATCACAGGATACACTTCCACAATTAATTAATAGT


ACTGAAGCAGAAATTAGAAATATTTTATATCAAGGACAAATTGGTAAGCAAAATAAACCAAGTGTAACTACA


CATGCTAAAGTTAGTGATCAAGAACTAGGTAAGCAGTCAAGACGTTCTCAAGATATCATTAAGTCATTAGGT


TTCCTTTCATCAGACCAAAAAGATATTTTAGTTAAATATATTAGCTCTTCAAAAGATTCGCAACTTATTCTT


AAATTTGTAACTCAAGCCACGCAACTGAATAATGCTGAATCAACAAAAGCTAAGCAAATGGCTCAAAATGAC


GTGGCCTTAATAAAAAATATAAGCCCCGAAGTCTTAGAAGAATATAAAGAAAAAATTCAAAGAGCTAGCACT


AAGAGTCAAGTTGATGAGTTTGTAGCAGAAGCTAAAAAAGTTGTTAATTCCAATAAAGAAACGTTGGTAAAT


CAGGCCAATGGTAAAAAGCAAGAAATTGATAAGTTAGAAAATTTATCTAACGATGAAATGTTGAGATATAAT


ACTGCAATTGATAATGTAGTGAAACAGTATAATGAAGGTAAGCTCAATATTACTGCTGCAATGAATGCTTTA


AATAGTATTAAGCAAGCAGCACAGGAAGTTGCCCAGAAAAACTTACAAAAGCAGTATGCTAAAAAAATTGAA


AGAATAAGTTCAAAAGGATTAGCGTTATCTAAAAAGGCTAAAGAAATTTATGAAAAGCATAAAAGTATTTTG


CCTACACCTGGATATTATGCAGACTCTGTGGGAACTTATTTGAATAGGTTTAGAGATAAACAAACTTTCGGA


AATAGGAGTGTTTGGACTGGTCAAAGTGGACTTGATGAAGCAAAAAAAATGCTTGATGAAGTCAAAAAGCTT


TTAAAAGAACTTCAAGACCTTACCAGAGGTACTAAAGAAGATAAAAAACCAGACGTTAAGCCAGAAGCCAAA


CCAGAGGCCAAACCAAATATTCAAGTACCTAAACAAGCACCTACAGAAGCTGCAAAACCAGCTTTGTCACCA


GAAGCCTTGACAAGATTGACTACATGGTATAATCAAGCTAAAGATCTGCTTAAAGATGATCAAGTAAAGGAC


AAATACGTAGATATACTTGCAGTTCAAAAAGCTGTTGACCAAGCTTATGATCATGTGGAAGAGGGAAAATTT


ATTACCACTGATCAAGCAAATCAATTAGCTAACAAGCTACGTGATGCTTTACAAAGTTTAGAATTAAAAGAT


AAAAAAGTAGCCAAACCAGAAGCCAAACCAGAGGCCAAACCAGAAGCTAAGCCAGAAGCTAAGCCAGAAGCT


AAGCCAGAAGCTAAGCCAGAGGCCAAACCAGAAGCTAAGCCAGACGTTAAGCCAGAAGCTAAACCAGACGTT


AAACCAGAGGCTAAGCCAGAAGCTAAACCAGAGGCTAAGTCAGAAGCTAAACCAGAGGCTAAGCTAGAAGCT


AAACCAGAGGCCAAACCAGCAACCAAAAAATCGGTTAATACTAGCGGAAACTTGGCGGCTAAAAAAGCTATT


GAAAACAAAAAGTATAGTAAAAAATTACCATCAACGGGTGAAGCCGCAAGTCCACTCTTAGCAATTGTATCA


CTAATTGTTATGTTAAGTGCAGGTCTTATTACGATAGTTTTAAAGCATAAAAAAAATTAA


 


ORF amino acid sequence (SEQ ID NO: 179):


MNNNEKKVKYFLRKTAYGLASMSAAFAVCSGIVHADTSSGISASIPHKKQVNLGAVTLKNLISKYRGNDKAI


AILLSRVNDFNRASQDTLPQLINSTEAEIRNILYQGQIGKQNKPSVTTHAKVSDQELGKQSRRSQDIIKSLG


FLSSDQKDILVKYISSSKDSQLILKFVTQATQLNNAESTKAKQMAQNDVALIKNISPEVLEEYKEKIQRAST


KSQVDEFVAEAKKVVNSNKETLVNQANGKKQEIDKLENLSNDEMLRYNTAIDNVVKQYNEGKLNITAAMNAL


NSIKQAAQEVAQKNLQKQYAKKIERISSKGLALSKKAKEIYEKHKSILPTPGYYADSVGTYLNRFRDKQTFG


NRSVWTGQSGLDEAKKMLDEVKKLLKELQDLTRGTKEDKKPDVKPEAKPEAKPNIQVPKQAPTEAAKPALSP


EALTRLTTWYNQAKDLLKDDQVKDKYVDILAVQKAVDQAYDHVEEGKFITTDQANQLANKLRDALQSLELKD


KKVAKPEAKPEAKPEAKPEAKPEAKPEAKPEAKPEAKPDVKPEAKPDVKPEAKPEAKPEAKSEAKPEAKLEA


KPEAKPATKKSVNTSGNLAAKKAIENKKYSKKLPSTGEAASPLLAIVSLIVMLSAGLITIVLKHKKN
















TABLE 13





(A and B): Overview over the two runs of sequencing of the six antigens from


various GBS strains; SEQ ID NOs of the corresponding proteins are listed.







A











Strain name
Serotype
gbs0233
gbs1087
gbs1309





IC97(III)
III
SEQ ID NO: 235
n.d.
n.d.


IC98(II)
II
SEQ ID NO: 236
SEQ ID NO: 287
SEQ ID NO: 317


IC105(IV)
IV
SEQ ID NO: 59
SEQ ID NO: 71
SEQ ID NO: 83


IC108(III)
III
SEQ ID NO: 237
SEQ ID NO: 288
n.d.


IC216(Ib)
Ib
SEQ ID NO: 238
SEQ ID NO: 289
SEQ ID NO: 318


IC244(III)
III
SEQ ID NO: 239
n.d.
SEQ ID NO: 319


IC245(Ib)
Ib
SEQ ID NO: 240
SEQ ID NO: 290
SEQ ID NO: 320


IC246(III)
III
SEQ ID NO: 241
SEQ ID NO: 291
n.d.


IC247(III)
III
SEQ ID NO: 242
n.d.
SEQ ID NO: 321


IC250(Ib)
Ib
SEQ ID NO: 243
SEQ ID NO: 292
SEQ ID NO: 322


IC251(V)
V
SEQ ID NO: 244
n.d.
SEQ ID NO: 323


IC252(III)
III
SEQ ID NO: 245
SEQ ID NO: 293
SEQ ID NO: 324


IC252/2(III)
III
n.d.
n.d.
n.d.


IC253(III)
III
SEQ ID NO: 246
SEQ ID NO: 294
SEQ ID NO: 325


IC254(II)
II
SEQ ID NO: 247
n.d.
n.d.


IC255(V)
V
SEQ ID NO: 248
SEQ ID NO: 295
SEQ ID NO: 326


IC287(V)
V
SEQ ID NO: 249
n.d.
SEQ ID NO: 327


IC288(Ia)
Ia
SEQ ID NO: 250
n.d.
SEQ ID NO: 328


IC289(Ib)
Ib
SEQ ID NO: 251
SEQ ID NO: 296
SEQ ID NO: 329


IC290(III)
III
SEQ ID NO: 252
SEQ ID NO: 297
SEQ ID NO: 330


IC291(V)
V
SEQ ID NO: 253
SEQ ID NO: 298
SEQ ID NO: 331


IC304(V)
V
SEQ ID NO: 254
SEQ ID NO: 299
SEQ ID NO: 332


IC305(II)
II
SEQ ID NO: 255
SEQ ID NO: 300
SEQ ID NO: 333


IC306(Ib)
Ib
SEQ ID NO: 256
SEQ ID NO: 301
SEQ ID NO: 334


IC361(Ib)
Ib
SEQ ID NO: 257
SEQ ID NO: 302
SEQ ID NO: 335


IC363(III)
III
SEQ ID NO: 258
n.d.
SEQ ID NO: 336


IC364(III)
III
SEQ ID NO: 259
SEQ ID NO: 303
SEQ ID NO: 337


IC365(Ia)
Ia
SEQ ID NO: 260
SEQ ID NO: 304
SEQ ID NO: 338


IC366(n.t.)
non
SEQ ID NO: 261
n.d.
SEQ ID NO: 339



typeable


IC367(II)
II
SEQ ID NO: 262
n.d.
n.d.


IC368(Ia)
Ia
SEQ ID NO: 263
SEQ ID NO: 305
SEQ ID NO: 340


IC377(V)
V
SEQ ID NO: 264
SEQ ID NO: 306
SEQ ID NO: 341


IC379(Ib)
Ib
SEQ ID NO: 265
SEQ ID NO: 307
SEQ ID NO: 342


IC432(Ib)
Ib
SEQ ID NO: 266
n.d.
SEQ ID NO: 343


IC434(III)
III
SEQ ID NO: 267
SEQ ID NO: 308
n.d.


IC455(III)
III
SEQ ID NO: 268
n.d.
SEQ ID NO: 344


IC457(II)
II
SEQ ID NO: 269
SEQ ID NO: 309
SEQ ID NO: 345


IC458(Ia)
Ia
SEQ ID NO: 60
SEQ ID NO: 72
SEQ ID NO: 84


IC459(Ib)
Ib
SEQ ID NO: 270
n.d.
SEQ ID NO: 346


IC460(II)
II
SEQ ID NO: 271
n.d.
SEQ ID NO: 347


IC461(Ib)
Ib
SEQ ID NO: 272
SEQ ID NO: 310
SEQ ID NO: 348


IC462(II)
II
SEQ ID NO: 273
n.d.
n.d.


IC463(Ib)
Ib
SEQ ID NO: 274
n.d.
SEQ ID NO: 349


IC469(V)
V
SEQ ID NO: 275
SEQ ID NO: 311
SEQ ID NO: 350


IC470(V)
V
SEQ ID NO: 276
n.d.
n.d.


126H4A(Ia)
Ia
SEQ ID NO: 277
n.d.
SEQ ID NO: 351


5095S2(Ib)
Ib
SEQ ID NO: 278
n.d.
n.d.


6313(III)
III
SEQ ID NO: 279
SEQ ID NO: 230
SEQ ID NO: 352


12351(IV)
IV
SEQ ID NO: 280
n.d.
SEQ ID NO: 353


NEM316,
III
SEQ ID NO: 229
n.d.
SEQ ID NO: 231


12403(III)


12403/2(III)
III
SEQ ID NO: 281
n.d.
SEQ ID NO: 354


12401(Ib)
Ib
SEQ ID NO: 56
SEQ ID NO: 68
SEQ ID NO: 80


COH1(III)
III
SEQ ID NO: 58
SEQ ID NO: 70
SEQ ID NO: 82


BAA23(V)
V
SEQ ID NO: 57
SEQ ID NO: 69
SEQ ID NO: 81


0176H4A(II)
II
SEQ ID NO: 55
SEQ ID NO: 67
SEQ ID NO: 79


A909(Ia/c)
Ia/c
SEQ ID NO: 282
SEQ ID NO: 312
SEQ ID NO: 355


C388/90(Ia/c)
Ia/c
SEQ ID NO: 283
SEQ ID NO: 313
SEQ ID NO: 356


BAA22(III)
III
SEQ ID NO: 284
SEQ ID NO: 314
SEQ ID NO: 357


2603V/R(V)
V
SEQ ID NO: 285
SEQ ID NO: 315
SEQ ID NO: 358


49447(V)
V
SEQ ID NO: 286
SEQ ID NO: 316
SEQ ID NO: 359


BAA611(V)
V
n.d.
n.d.
n.d.










B











Strain name
Serotype
gbs1477
gbs1478
gbs2018





IC97(III)
III
SEQ ID NO: 101
n.d.
SEQ ID NO: 379


IC98(II)
II
SEQ ID NO: 102
SEQ ID NO: 187
SEQ ID NO: 380


IC105(IV)
IV
SEQ ID NO: 103
SEQ ID NO: 188
SEQ ID NO: 178


IC108(III)
III
n.d.
SEQ ID NO: 189
n.d.


IC216(Ib)
Ib
SEQ ID NO: 104
SEQ ID NO: 190
SEQ ID NO: 381


IC244(III)
III
n.d.
SEQ ID NO: 191
SEQ ID NO: 382


IC245(Ib)
Ib
SEQ ID NO: 105
SEQ ID NO: 192
SEQ ID NO: 383


IC246(III)
III
n.d.
SEQ ID NO: 193
n.d.


IC247(III)
III
n.d.
SEQ ID NO: 194
SEQ ID NO: 384


IC250(Ib)
Ib
SEQ ID NO: 106
SEQ ID NO: 195
SEQ ID NO: 385


IC251(V)
V
SEQ ID NO: 107
SEQ ID NO: 196
SEQ ID NO: 386


IC252(III)
III
SEQ ID NO: 108
n.d.
SEQ ID NO: 387


IC252/2(III)
III
SEQ ID NO: 360
n.d.
n.d.


IC253(III)
III
SEQ ID NO: 109
SEQ ID NO: 197
SEQ ID NO: 388


IC254(II)
II
SEQ ID NO: 110
n.d.
SEQ ID NO: 389


IC255(V)
V
SEQ ID NO: 111
n.d.
SEQ ID NO: 390


IC287(V)
V
SEQ ID NO: 112
n.d.
SEQ ID NO: 391


IC288(Ia)
Ia
n.d.
n.d.
n.d.


IC289(Ib)
Ib
SEQ ID NO: 113
SEQ ID NO: 198
SEQ ID NO: 392


IC290(III)
III
n.d.
n.d.
SEQ ID NO: 393


IC291(V)
V
SEQ ID NO: 114
SEQ ID NO: 199
SEQ ID NO: 394


IC304(V)
V
SEQ ID NO: 115
SEQ ID NO: 200
SEQ ID NO: 395


IC305(II)
II
SEQ ID NO: 116
SEQ ID NO: 201
SEQ ID NO: 396


IC306(Ib)
Ib
SEQ ID NO: 117
SEQ ID NO: 202
SEQ ID NO: 397


IC361(Ib)
Ib
SEQ ID NO: 118
n.d.
SEQ ID NO: 398


IC363(III)
III
SEQ ID NO: 119
n.d.
SEQ ID NO: 399


IC364(III)
III
n.d.
n.d.
n.d.


IC365(Ia)
Ia
SEQ ID NO: 120
SEQ ID NO: 363
SEQ ID NO: 400


IC366(n.t.)
non
n.d.
SEQ ID NO: 364
SEQ ID NO: 401



typeable


IC367(II)
II
SEQ ID NO: 121
SEQ ID NO: 365
SEQ ID NO: 402


IC368(Ia)
Ia
n.d.
n.d.
SEQ ID NO: 403


IC377(V)
V
SEQ ID NO: 122
SEQ ID NO: 366
SEQ ID NO: 404


IC379(Ib)
Ib
SEQ ID NO: 123
SEQ ID NO: 367
SEQ ID NO: 405


IC432(Ib)
Ib
SEQ ID NO: 124
SEQ ID NO: 368
SEQ ID NO: 406


IC434(III)
III
n.d.
n.d.
n.d.


IC455(III)
III
SEQ ID NO: 125
SEQ ID NO: 369
SEQ ID NO: 407


IC457(II)
II
SEQ ID NO: 126
SEQ ID NO: 370
SEQ ID NO: 408


IC458(Ia)
Ia
SEQ ID NO: 127
SEQ ID NO: 203
SEQ ID NO: 179


IC459(Ib)
Ib
SEQ ID NO: 128
SEQ ID NO: 371
SEQ ID NO: 409


IC460(II)
II
SEQ ID NO: 129
SEQ ID NO: 372
SEQ ID NO: 410


IC461(Ib)
Ib
SEQ ID NO: 130
SEQ ID NO: 373
SEQ ID NO: 411


IC462(II)
II
SEQ ID NO: 131
SEQ ID NO: 374
SEQ ID NO: 412


IC463(Ib)
Ib
n.d.
n.d.
SEQ ID NO: 413


IC469(V)
V
n.d.
SEQ ID NO: 375
SEQ ID NO: 414


IC470(V)
V
SEQ ID NO: 132
SEQ ID NO: 376
SEQ ID NO: 415


126H4A(Ia)
Ia
SEQ ID NO: 94
n.d.
SEQ ID NO: 416


5095S2(Ib)
Ib
SEQ ID NO: 96
n.d.
SEQ ID NO: 417


6313(III)
III
SEQ ID NO: 97,
SEQ ID NO: 233
SEQ ID NO: 418




SEQ ID NO: 232


12351(IV)
IV
SEQ ID NO: 92
n.d.
SEQ ID NO: 419


NEM316,
III
n.d.
n.d.
SEQ ID NO: 234


12403(III)


12403/2(III)
III
SEQ ID NO: 361
n.d.
SEQ ID NO: 420


12401(Ib)
Ib
SEQ ID NO: 93
SEQ ID NO: 185
SEQ ID NO: 176


COH1(III)
III
n.d.
n.d.
SEQ ID NO: 421


BAA23(V)
V
SEQ ID NO: 98
SEQ ID NO: 186
SEQ ID NO: 177


0176H4A(II)
II
SEQ ID NO: 91
SEQ ID NO: 377
SEQ ID NO: 175


A909(Ia/c)
Ia/c
n.d.
n.d.
n.d.


C388/90(Ia/c)
Ia/c
SEQ ID NO: 100
n.d.
SEQ ID NO: 422


BAA22(III)
III
n.d.
n.d.
SEQ ID NO: 423


2603V/R(V)
V
SEQ ID NO: 362
SEQ ID NO: 378
SEQ ID NO: 424


49447(V)
V
SEQ ID NO: 95
n.d.
SEQ ID NO: 425


BAA611(V)
V
SEQ ID NO: 99
n.d.
n.d.


















MEGA

## Claims

1. A composition comprising a first protective protein and a second protective protein,
i) wherein the first protective protein consists of the amino acid sequence of SEQ ID NO: 5 (gbs1478p) or functionally active protein that consists of at least 95% identity to the sequence of SEQ ID NO:5; and
wherein the functionally active protein shows at least 80% of the protection shown for the protective protein of SEQ ID NO: 5 against challenge with at least one strain of S. agalactiae used in a dose of 106-108 cfu in a lethal sepsis model, wherein the strain is selected from the group consisting of C388/90, A909, ATCC12401, ATCC1243, COH1, BAA22, BAA23, 2603V/R, and ATCC49447;
ii) wherein the second protective protein consists of the amino acid sequence of SEQ ID NO: 6 (gbs2018p) or functionally active protein that consists of at least 95% identity to the sequence of SEQ ID NO: 6; and
wherein the functionally active protein shows at least 80% of the protection shown for the protective protein of SEQ ID NO: 6 against challenge with at least one strain of S. agalactiae used in a dose of 106-108 cfu in a lethal sepsis model, wherein the strain is selected from the group consisting of C388/90, A909, ATCC12401, ATCC1243, COH1, BAA22, BAA23, 2603V/R, and ATCC49447.

2. The composition of claim 1, wherein the first protective protein consists of the amino acid sequence of SEQ ID NO: 5 (gbs1478p).

3. The composition of claim 1, wherein the second protective protein consists of the amino acid sequence of SEQ ID NO: 6 (gbs2018p).

4. The composition of claim 1, wherein
i) the first protective protein consists of the amino acid sequence of SEQ ID NO: 5 gbs1478p); and
ii) the second protective protein consists of the amino acid sequence of SEQ ID NO: 6 (gbs2018p).

5. The composition of claim 1, wherein the first protective protein and the second protective protein are combined into at least one fusion protein.

6. A pharmaceutical composition comprising
(i) the composition according to claim 1; and
(ii) optionally a pharmaceutically acceptable carrier or excipient.

7. The pharmaceutical composition of claim 6, further comprising an immunostimulatory substance.

8. The pharmaceutical composition of claim 7, wherein the immunostimulatory substance is an adjuvant.

9. A pharmaceutical composition comprising
(i) the composition according to claim 2; and
(ii) optionally a pharmaceutically acceptable carrier or excipient.

10. The pharmaceutical composition of claim 9, further comprising an immunostimulatory substance.

11. The pharmaceutical composition of claim 10, wherein the immunostimulatory substance is an adjuvant.

12. A method of inducing an immune response in a subject against S. agalactiae infection comprising administering an effective amount of the composition of claim 1 to said subject.