Abstract:
The invention is concerned with the identification of a novel class of bacterial polysaccharide biosynthetic operons and an ovel clas of regulatory operons involved with polysaccharide biosynthesis, bacterial attachment and biofilm development. Bacterial strains which possess a polysaccharide biosynthetic operon of the type provide by the invention are capable of producing polysaccharide wtih industrial implications. Bacterial strains which possess a regulatory operon of the type provided by the invention may be targeted by pharmaceutical/chemical agents to prevent bacterial attachment and biofilm development.

Description:
FIELD OF THE INVENTION  
         [0001]    The present invention is concerned with the identification of a novel class of bacterial polysaccharide biosynthetic operons and a novel class of regulatory operons involved with polysaccharide biosynthesis, bacterial attachment and biofilm development. Bacterial strains which possess a polysaccharide biosynthetic operon of the type provided by the invention are capable of producing a polysaccharide with industrial implications. Bacterial strains which possess a regulatory operon of the type provided by the invention may be targeted by pharmaceutical/chemical agents to prevent bacterial attachment and biofilm development.  
           [0002]    Also provided by the invention is a process for the production of the polysaccharide, methods of isolating/engineering polysaccharide-producing bacterial strains and also novel enzymes involved in polysaccharide biosynthesis, methods to screen chemical libraries for pharmaceutical/chemical agents to prevent bacterial attachment and biofilm development and also novel proteins involved in the regulation of polysaccharide synthesis, bacterial attachment and biofilm development.  
         BACKGROUND TO THE INVENTION  
         [0003]    Cellulose and modified celluloses are valuable polymers with a wide range of applications, for example, in the food, clothing, paint and paper-producing industries and also in medicine, particularly the production of artificial veins and wound dressings. Most cellulose is derived from plants, but there is increasing interest in bacterial-derived celluloses, because of their value in the production of speciality items, such as certain papers, natural thickening agents for the food industry and artificial tissues for use in medicine. The vast majority of industrial uses are, in fact, for modified celluloses. It is the substitution pattern on the primary cellulosic backbone that creates the various properties exhibited by industrially useful modified celluloses.  
           [0004]    Despite the obvious usefulness of bacterial cellulose, to date only a single cellulose-producing bacterium, Acetobacter, is exploited by industry (see general review by Ross, P. et al.  Microbiol. Rev.  1991, 55: 35-50). It is therefore an object of the present invention to provide an alternative source of industrially useful cellulose-like bacterial polysaccharides.  
           [0005]    The present inventors have identified a novel class of polysaccharide biosynthetic operon, the genetic organisation of which differs significantly from the cellulose biosynthetic operon of Acetobacter. Of key importance, the inventors have identified a second locus involved in the regulation of polysaccharide biosynthesis. Bacterial strains which possess the polysaccharide biosynthetic operon and in which enzyme-encoding genes of the operon are expressed, including certain strains of Pseudomonas and  E. coli , are capable of producing large amounts of a particular type of polysaccharide.  
           [0006]    The inventors have also determined that the second regulatory locus is involved in bacterial attachment. Bacterial attachment is a necessary first step in the development of biofilms. Isogenic pairs of bacterial strains, one of which lacks the regulatory locus, can provide a screen for chemical or pharmaceutical agents that block attachment and biofilm growth.  
         DESCRIPTION OF THE INVENTION  
         [0007]    Glucan-Like Polysaccharide  
           [0008]    In a first aspect the invention provides a glucan-like polysaccharide produced by an exopolysaccharide-producing bacterial strain, said bacterial strain being characterised in that it expresses one or more enzyme-encoding genes of a wss-like operon.  
           [0009]    The polysaccharide of the invention can be derived from any bacterial strain which expresses one or more enzyme-encoding genes of a wss-like operon and is usually produced as an extracellular polysaccharide (or exopolysaccharide). Bacterial strains which produce the polysaccharide of the invention may be referred to herein as “exopolysaccharide-producing bacterial strains”.  
           [0010]    The term “exopolysaccharide-producing bacterial strain” encompasses any bacterial strain which has a wss-like polysaccharide biosynthetic operon and in which one or more of the genes encoding subunits of cellulose synthase are expressed. Also encompassed within the scope of the term “exopolysaccharide-producing bacterial strain” are recombinant strains which have been engineered to express one or more enzyme-encoding genes of a wss-like operon and also strains which have a wss-like operon and have been engineered to over-express a regulator of the wss-like operon. The characteristics and construction of these recombinant strains will be more fully explained below. Finally, the term “exopolysaccharide-producing bacterial strain” also encompasses mutagenized strains derived from parent strains having a wss-like operon, for example strains wherein one or more genes of the wss-like operon which are not expressed in the parent strain are expressed in the mutagenized strain. The construction of such mutagenized strains will be described in more detail below.  
           [0011]    Different exopolysaccharide producing bacterial strains may produce glucan-like polysaccharides which are slightly different in terms of structure and/or chemical composition. It is to be understood that the term “glucan-like polysaccharide” does not refer to any one single substance but is rather a generic term which encompasses exopolysaccharides produced by a wide range of exopolysaccharide producing bacterial strains.  
           [0012]    In a preferred embodiment the exopolysaccharide-producing bacterial strain is an “evolved variant” of an ancestral strain, wherein the ancestral strain has a wss-like polysaccharide biosynthetic operon but does not naturally produce the glucan-like polysaccharide provided by the invention. As exemplified herein, the inventors have observed that a wild-type strain which possesses a wss-like operon can evolve into polysaccharide producing evolved variants when cultured in a novel environment, such as the static broth culture described herein. Exopolysaccharide-producing evolved variants of Pseudomonas sp. may be distinguished from the corresponding ancestral strain, for example by virtue of a characteristic “wrinkly spreader” (WS) morphology (described by Rainey &amp; Travisano, 1998 , Nature,  394: 69-72). Procedures for the isolation of exopolysaccharide-producing evolved variant strains will be described hereinbelow.  
           [0013]    “Wss-like operon” is a generic term used herein to describe a novel class of bacterial operons containing genes which encode enzymes involved in the biosynthesis of glucan-like polysaccharides. In general terms, wss-like operons are distinguished from the cellulose biosynthetic operons of Acetobacter because they lack a gene encoding the enzyme responsible for cellulose crystallization. Encompassed within the term “wss-like operon” are operons which are homologous to the wss operon of  Pseudomonas fluorescens,  the complete nucleotide sequence of which is given herein. Preferably, the wss-like operon comprises a sequence of nucleotides which shares at least 50% nucleotide sequence identity with the sequence of nucleotides shown in FIG. 2 (SEQ ID NO: 1) or FIG. 27 (SEQ ID NO: 26). Even more preferably, the wss-like operon may comprise a sequence of nucleotides at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 90% or at least 95% identical to the  P. fluorescens  wss operon shown in FIG. 2 (SEQ ID NO: 1) or the  E. coli  yhj operon shown in FIG. 27 (SEQ ID NO: 26).  
           [0014]    In accordance with the invention, percent identity of nucleotide and amino acid sequences may be calculated based on an optimal alignment of the sequences to be compared, taking account of nucleotide/amino acid insertions and deletions. An optimal alignment can be assembled using the BLAST algorithm which is well known in the art.  
           [0015]    Percentage nucleotide identity may be calculated by comparing entire wss or yhj operon sequences or by comparing the coding regions of the individual genes of the operons. In the latter case, the coding regions of homologous genes should be compared. A value for the overall percentage sequence identity may then be derived by taking an average over all the individual homologous coding regions. A complete annotation of the  P. fluorescens  wss operon, showing the positions of the coding regions is listed elsewhere in this specification. Similarly, the positions of the individual coding regions within the  E. coli  yhj operon are given below.  
           [0016]    As well as having a significant degree of nucleotide sequence identity with the nucleotide sequences illustrated in FIG. 2 (SEQ ID NO: 1) or FIG. 27 (SEQ ID NO: 26), the wss-like operon may or may not share substantial organisational similarity with the  Pseudomonas fluorescens  wss and  E. coli  yhj operons, meaning that the homologous genes are arranged in the same order within the operon. “Substantial organisational similarity” with the  P. fluorescens  wss operon should be taken to mean that at least the genes encoding the subunits of cellulose synthase should be arranged in the same order as they are in the  Pseudomonas fluorescens  wss operon. As illustrated in FIG. 1, the genes encoding enzyme subunits which are homologous to cellulose synthase subunits from other bacterial species are arranged 5′-wssB-wssC-wssE-3′. These genes and the enzymes they encode may be designated herein as “cellulose synthases” but this is on the basis of homology to cellulose synthase genes from other bacterial species. The use of this nomenclature should not be taken to imply that the final polysaccharide products synthesised by the action of these enzymes are pure celluloses (i.e. pure 1-4 β-linked glucan) or that the activity of these enzymes is limited to the synthesis of pure celluloses. Wss-like operons may be distinguished from known bacterial cellulose biosynthetic operons (e.g. the acetobacter Bcs operon) by the absence of the gene responsible for cellulose crystallization and the presence of additional genes (wss G H I; alg F I J) whose enzyme products are involved in acetylation of polysaccharides. Although it is possible that the enzymes encoded by the wssA-E genes may produce a product which is a substantially pure cellulose, the enzymes encoded by wss G H I might then modify this product.  
           [0017]    The term “enzyme-encoding gene” as used herein refers to a gene which encodes a protein product which functions as an enzyme or as an enzyme subunit.  
           [0018]    Knowledge of the primary nucleotide sequence and the structural organisation of the  P. fluorescens  wss and  E. coli  yhj operons enables the identification of other microorganisms which have a wss-like operon. This can be accomplished using a variety of techniques which are known in the art. In order to find out whether a given microorganism has a wss-like operon, a labelled nucleic acid probe corresponding to a part of the wss operon could be used to probe a Southern blot of genomic DNA. Genomic DNA fragments containing the wss-like operon, or parts thereof, could then be isolated by probing a library of genomic DNA from the organism, e.g. a library of bacterial chromosomal DNA fragments. Preferably, the labelled probe fragment would correspond to a region of one of the enzyme-encoding genes of the wss operon which is highly conserved cross-species. Procedures for the preparation of suitable labelled probe fragments, Southern blotting, construction of chromosomal libraries, cross-species library screening, recovery of positive clones and sequencing of the DNA inserts would be well known to one skilled in the art (see, for example, Sambrook, Fritsch &amp; Maniatis,  Molecular Cloning: A Laboratory Manual,  Second Edition (1989) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.; F. M. Ausubel et al. (eds.),  Current Protocols in Molecular Biology,  John Wiley &amp; Sons, Inc. (1994)).  
           [0019]    As an alternative to the library screening approach, oligonucleotide primers corresponding to suitable regions of the  P. fluorescens  wss operon or the  E. coli  yhj operon could be used for PCR amplification of homologous wss operon sequences from DNA isolated from other bacterial species. Again, standard procedures for purification of chromosomal DNA, PCR amplification and cloning and sequencing of PCR products are well known in the art (Sambrook, et al. supra; Ausubel, et al. supra).  
           [0020]    Finally, a vast amount of publicly available nucleotide sequence data from a wide range of different organisms is accessible electronically via the Internet, see for example, www.ncbi.nlm.nih.gov. Therefore, one approach to identifying wss-like operons in other species would be a “bioinformatics” approach using the nucleotide sequence of the  P. fluorescens  wss operon or the  E. coli  yhj operon or a fragment thereof to perform a search of the available databases.  
           [0021]    As will be illustrated in the Examples included herein, the glucan-like polysaccharide of the invention is obtainable by,  
           [0022]    (a) culturing an exopolysaccharide-producing bacterial strain;  
           [0023]    (b) lysing the bacteria overnight at 37° C. in a lysis solution of 20 mM Tris.HCl pH8.0, 5 mM MgCl 2 , 0.5% Sarkosyl, 1 mg/ml fresh lysozyme;  
           [0024]    (c) treating the lysed sample obtained in step (b) with DNase and RNase; and  
           [0025]    (d) incubating the sample obtained in step (c) for 24 hr with a second lysis solution of 500 mM EDTA pH 9.0, 1% Sarkosyl, 1.5 mg/ml Proteinase K.  
           [0026]    However, it is to be understood that the invention is in no way limited to polysaccharide produced according to the process steps listed as steps (a) to (d) above.  
           [0027]    The polysaccharide provided by the invention may also be identified on the basis of positive staining with a staining reagent which specifically stains polysaccharides having a predominantly β-linked glucan structure, for example calcofluor.  
           [0028]    In terms of chemical structure, the polysaccharide of the invention is defined as a polymer having glucose residues as the main constituent of the polymeric backbone, wherein the glucose residues are linked by glycosidic bonds, which may be of either α or β configuration and may be of any of the following linkage varieties; 1-3,1-4 or 1-6. In addition, the polymer backbone may have integrated into its structure other hexose sugar residues or derivatised hexose residues, for example galactose, mannose or galacturonic acid. The residues in the backbone may further be substituted at any position by hexose or pentose residues, derivatised hexose or pentose residues or other functional groups including, but not limited to, acidic functional groups such as acetyl groups. The polymer may be of any degree of polymerisation.  
           [0029]    The polysaccharide of the invention may be referred to herein as being a “glucan-like” polymer, since its structure is based predominantly, though not exclusively, on a β-linked glucan backbone that may be substituted with, for example, other sugars and also functional groups. It is to be understood that the polysaccharide of the invention is not a pure cellulose, i.e. unsubstituted, since it is generally observed to be amorphous in structure rather than crystalline.  
           [0030]    The polysaccharide polymers provided by the invention may have a plurality of structures and associated properties. These properties may include, but not exclusively, degree of crystallinity, degree of polymerisation, degree, extent and pattern of substitution, ability to form fluids of varying viscosity when the polymers are integrated into a fluid substance, ability to control enzymic reaction rates when in an environment with active enzyme components, ability to control or otherwise alter the tensile strength of a substance when integrated or mixed with such a substance or product, ability to control or otherwise alter the motility of bacteria in an environment into which the polymer has been introduced, ability to control or otherwise alter the attachment of bacteria to surfaces in an environment into which the polymer has been introduced ability to control or otherwise alter an organisms ability to metabolise any substance when the polymer is in the environment of the substance.  
           [0031]    Methods of Isolating Exopolysaccharide-Producing Bacterial Strains  
           [0032]    In a still further aspect, the invention provides a method of isolating an exopolysaccharide-producing bacterial strain, which method comprises the steps of:  
           [0033]    (a) growing a wild type bacterial strain, the genome of which comprises a wss-like operon, in a static broth culture;  
           [0034]    (b) isolating a variant strain having wrinkly spreader morphology;  
           [0035]    (c) optionally, screening colonies of the wrinkly spreader morph obtained in step (b) for the production of polysaccharide.  
           [0036]    The above method of the invention can be used to isolate exopolysaccharide-producing variants of any wild type bacterial strain having a wss-like operon. This bacterial strain is commonly referred to herein as an “ancestral” strain. Advantageously, the method of the invention can be used to isolate exopolysaccharide-producing strains of Pseudomonas and  E. coli . Preferred ancestral Pseudomonas strains which can be used in the method of the invention include  Pseudomonas fluorescens  SBW25. Preferred  E. coli  ancestral strains which can be used in the method of the invention include  E.coli  K12.  
           [0037]    Ancestral  P. fluorescens  SBW25 shows no evidence of cellulose or modified cellulose production in vitro (although available data suggests that the wss operon is active when the bacterium is associated with plant surfaces). Mutant forms which were later shown to produce a glucan-like polysaccharide were obtained following selection of the ancestral (smooth; SM) genotype in stationary broth vials over the course of 10 days (see Rainey &amp; Travisano, Nature, 394, 69-72). Among a diverse collection of mutant genotypes that evolve are a class known as wrinkly spreaders (WS). The name of this class of mutants reflects their morphology on agar plates and in static broth microcosms they colonise the interface between liquid and air. Subsequent genetic analysis of WS morphology revealed the wss operon, the wsp operon and various other genes described in this application with a role in the production of the glucan-like polysaccharide.  
           [0038]    The distribution of the wss operon among other Pseudomonas strains has been examined using gene probes made from the  P. fluorescens  SBW25 wss operon. Positive signals can be observed in a number of strains. The sequence of the wss genes can therefore be used to identify other glucan-like polysaccharide-producing bacteria. The inventors have also used the evolutionary selection procedure to show that strains with wss genes are capable of generating the WS phenotype. Thus evolutionary selection experiments also have the potential to reveal a latent ability to produce a glucan-like polysaccharide in a diverse range of Pseudomonas strains.  
           [0039]    The invention further provides an exopolysaccharide-producing bacterial strain which is obtainable by the method of the invention and the glucan-like polysaccharide produced by such a bacterial strain.  
           [0040]    In a still further aspect, the invention also provides a method of isolating an exopolysaccharide-producing bacterial strain which comprises the steps of exposing a bacterial strain, the genome of which comprises a wss-like operon, to a chemical mutagen; and identifying a mutant which produces a polysaccharide according to the invention.  
           [0041]    The chemical mutagen can be any mutagenic agent known in the art, preferably an agent which is known for use in random mutagenesis of bacterial chromosomes or a mixture of such agents.  
           [0042]    In one embodiment, the step of identifying a mutant which produces polysaccharide can be accomplished by plating out a large number of mutant colonies and staining with chemical stain specific for the polysaccharide (e.g. calcofluor, Congo red). Alternatively, exopolysaccharide-producing mutants can be identified by looking for variants having the wrinkly spreader colony morphology. This is a preferred method of identifying mutant strains of Pseudomonas. Following exposure to a chemical mutagen, the mutagenized bacteria may optionally be grown under culture conditions which favour the growth of exopolysaccharide-producing variants, for example the static culture conditions described herein (see Example 1).  
           [0043]    Also provided by the invention are exopolysaccharide-producing evolved variant and mutant strains which are obtainable according to the above-described methods, and the glucan-like polysaccharide produced by such strains.  
           [0044]    Novel Genes and Proteins  
           [0045]    The inventors have further identified a number of novel genes which encode components of the polysaccharide biosynthetic pathway of  P. fluorescens.    
           [0046]    Therefore, according to a further aspect of the invention there is provided an isolated nucleic acid molecule comprising the sequence of nucleotides from position 2200 to 18000 of the nucleotide sequence illustrated in FIG. 2 (SEQ ID NO: 1).  
           [0047]    [0047]FIG. 2 (SEQ ID NO: 1) shows the nucleotide sequence for a contiguous piece of DNA of 20,306 bp from the genome of  Pseudomonas fluorescens  SBW25. The wss operon, encoding genes homologous to known cellulose biosynthetic genes from other bacterial species and also associated genes, is located approximately between 2,200-18,000 bp. The operon consists of ten genes, designated wssA-J. A schematic figure showing the arrangement of the operon is shown in the accompanying FIG. 1.  
           [0048]    Isolated individual genes from the wss operon are also encompassed within the scope of the invention. Thus, the invention provides an isolated nucleic acid molecule encoding a WssA protein, said protein comprising the sequence of amino acids from position 145 to position 344 of the amino acid sequence illustrated in FIG. 3 (SEQ ID NO: 2) or from position 2 to position 344 of the amino acid sequence illustrated in FIG. 3 (SEQ ID NO: 2). Preferably the nucleic acid molecule comprises the sequence of nucleotides from position 2876 to 3478 or from position 2444 to 3478 of the nucleic acid sequence illustrated in FIG. 2 (SEQ ID NO: 1).  
           [0049]    Also provided by this aspect of the invention is a WssA protein comprising the sequence of amino acids from position 2 to 344 of the amino acid sequence illustrated in FIG. 3 (SEQ ID NO: 2) or from position 145 to position 344 of the amino acid sequence illustrated in FIG. 3 (SEQ ID NO: 2). The invention further provides a WssA protein encoded by a nucleic acid molecule according to the invention.  
           [0050]    The amino acid sequence shown in FIG. 3 (SEQ ID NO: 2) is the predicted translation of the longest open reading frame of the  Pseudomonas fluorescens  wssA gene. Translation of the WssA protein in vivo is expected to initiate at the first possible in-frame methionine codon (amino acid residue number 145 in the wssA predicted translated). However, it is possible that translation may initiate at the first in-frame valine codon (amino acid residue number 1 in the wssA translated sequence). In this case, the initiating amino acid of the protein product would still be methionine, not valine. Hence, the invention also provides a WssA protein comprising the amino acid sequence from position 2 to position 344 of the amino acid sequence illustrated in FIG. 3 (SEQ ID NO: 2) but having an additional N-terminal methionine residue.  
           [0051]    The invention further provides an isolated nucleic acid molecule encoding a WssB protein (sharing homology with cellulose synthase subunit A), said protein comprising the sequence of amino acids illustrated in FIG. 4 (SEQ ID NO: 3). Preferably the nucleic acid molecule comprises the sequence of nucleotides from position 3475 to 5694 of the nucleic acid sequence illustrated in FIG. 2 (SEQ ID NO: 1).  
           [0052]    Also provided by this aspect of the invention is a WssB protein comprising the sequence of amino acids illustrated in FIG. 4 (SEQ ID NO: 3). The invention further provides a WssB protein encoded by a nucleic acid molecule according to the invention.  
           [0053]    The amino acid sequence shown in FIG. 4 (SEQ ID NO: 3) is the predicted translation of part of the longest open reading frame of the Pseudomonas fluorescens wssB gene. Translation of the WssB protein in vivo is expected to initiate at the first possible in-frame methionine codon.  
           [0054]    The invention further provides an isolated nucleic acid molecule encoding a WssC protein (sharing homology with cellulose synthase subunit B), said protein comprising the sequence of amino acids from position 89 to position 689 of the amino acid sequence illustrated in FIG. 5 (SEQ ID NO: 4) or from position 2 to position 689 of the amino acid sequence illustrated in FIG. 5 (SEQ ID NO: 4). Preferably the nucleic acid molecule comprises the sequence of nucleotides from position 5884 to 7953 or from position 6148 to 7953 of the nucleic acid sequence illustrated in FIG. 2 (SEQ ID NO: 1).  
           [0055]    Also provided by this aspect of the invention is a WssC protein comprising the sequence of amino acids from position 89 to position 689 of the amino acid sequence illustrated in FIG. 5 (SEQ ID NO: 4) or from position 2 to position 689 of the amino acid sequence illustrated in FIG. 5 (SEQ ID NO: 4). The invention further provides a WssC protein encoded by a nucleic acid molecule according to the invention.  
           [0056]    The amino acid sequence shown in FIG. 5 (SEQ ID NO: 4) is the predicted translation of part of the longest open reading frame of the  Pseudomonas fluorescens  wssC gene. Translation of the WssC protein in vivo is expected to initiate at the first possible in-frame methionine codon (amino acid residue number 89 in the wssC translated sequence). However, it is possible that translation may initiate at the first in-frame valine codon (amino acid residue number 1 in the wssC translated sequence). In this case, the initiating amino acid of the protein product would still be methionine, not valine. Hence, the invention also provides a WssC protein comprising the amino acid sequence from position 2 to position 692 of the amino acid sequence illustrated in FIG. 5 (SEQ ID NO: 4) but having an additional N-terminal methionine residue.  
           [0057]    According to a further aspect of the invention there is provided an isolated nucleic acid molecule encoding a WssD protein (sharing homology with D-family cellulase associated with cellulose synthases), said protein comprising the sequence of amino acids from position 39 to position 436 of the amino acid sequence illustrated in FIG. 6 (SEQ ID NO: 5) or from position 2 to position 436 of the amino acid sequence illustrated in FIG. 6 (SEQ ID NO: 5). Preferably the nucleic acid molecule comprises the sequence of nucleotides from position 7884 to 9146 or from position 7950 to 9146 of the nucleic acid sequence illustrated in FIG. 2 (SEQ ID NO: 1).  
           [0058]    Also provided by this aspect of the invention is a WssD protein comprising the sequence of amino acids from position 39 to position 436 of the amino acid sequence illustrated in FIG. 6 (SEQ ID NO: 5) or from position 2 to position 436 of the amino acid sequence illustrated in FIG. 6 (SEQ ID NO: 5). The invention further provides a WssD protein encoded by a nucleic acid molecule according to the invention.  
           [0059]    The amino acid sequence shown in FIG. 6 (SEQ ID NO: 5) is the predicted translation of part of the longest open reading frame of the  Pseudomonas fluorescens  wssD gene. Translation of the WssD protein in vivo is expected to initiate at the first possible in-frame methionine codon (amino acid residue number 39 in the wssD translated sequence). However, it is possible that translation may initiate at the first in-frame valine codon (amino acid residue number 1 in the wssD translated sequence). In this case, the initiating amino acid of the protein product would still be methionine, not valine. Hence, the invention also provides a WssD protein comprising the amino acid sequence from position 2 to position 436 of the amino acid sequence illustrated in FIG. 6 (SEQ ID NO: 5) but having an additional N-terminal methionine residue.  
           [0060]    According to a further aspect of the invention there is provided an isolated nucleic acid molecule encoding a WssE protein (sharing homology with cellulose synthase subunit C), said protein comprising the sequence of amino acids illustrated in FIG. 7 (SEQ ID NO: 6). Preferably the nucleic acid molecule comprises the sequence of nucleotides from position 9128 to 12967 of the nucleic acid sequence illustrated in FIG. 2 (SEQ ID NO: 1).  
           [0061]    Also provided by this aspect of the invention is a WssE protein comprising the sequence of amino acids illustrated in FIG. 7 (SEQ ID NO; 6). The invention further provides a WssE protein encoded by a nucleic acid molecule according to the invention.  
           [0062]    The amino acid sequence shown in FIG. 7 (SEQ ID NO: 6) is a predicted translation of part of the longest open reading frame of the  Pseudomonas fluorescens  wssE gene. Translation of the WssE protein in vivo is expected to initiate at the first possible in-frame methionine codon (amino acid residue number 1 in the wssE translated sequence).  
           [0063]    The invention still further provides an isolated nucleic acid molecule encoding a WssF protein, said protein comprising the sequence of amino acids illustrated in FIG. 8 (SEQ ID NO: 7). Preferably the nucleic acid molecule comprises the sequence of nucleotides from position 12984 to 13649 of the nucleic acid sequence illustrated in FIG. 2 (SEQ ID NO: 1).  
           [0064]    Also provided by this aspect of the invention is an isolated WssF protein comprising the sequence of amino acids illustrated in FIG. 8 (SEQ ID NO: 7). The invention further provides a WssF protein encoded by a nucleic acid molecule according to the invention.  
           [0065]    The amino acid sequence shown in FIG. 8 (SEQ ID NO: 7) is a predicted translation of part of the longest open reading frame of the  Pseudomonas fluorescens  wssF gene. Translation of the WssF protein in vivo is expected to initiate at the first possible in-frame methionine codon (amino acid residue number 1 in the wssF translated sequence).  
           [0066]    The invention further provides an isolated nucleic acid molecule encoding a WssG protein, said protein comprising the sequence of amino acids illustrated in FIG. 9 (SEQ ID NO: 8). Preferably the nucleic acid molecule comprises the sequence of nucleotides from position 13649 to 14314 of the nucleic acid sequence illustrated in FIG. 2 (SEQ ID NO: 1).  
           [0067]    Also provided by this aspect of the invention is an isolated WssG protein comprising the sequence of amino acids illustrated in FIG. 9 (SEQ ID NO: 8). The invention further provides a WssG protein encoded by a nucleic acid molecule according to the invention.  
           [0068]    The amino acid sequence shown in FIG. 9 (SEQ ID NO: 8) is a predicted translation of part of the longest open reading frame of the  Pseudomonas fluorescens  wssG gene. Translation of the WssG protein in vivo is expected to initiate at the first possible in-frame methionine codon (amino acid residue number 1 in the wssG translated sequence).  
           [0069]    The invention further provides an isolated nucleic acid molecule encoding a WssH protein, said protein comprising the sequence of amino acids illustrated in FIG. 10 (SEQ ID NO: 9). Preferably the nucleic acid molecule comprises the sequence of nucleotides from position 14332 to 15738 of the nucleic acid sequence illustrated in FIG. 2 (SEQ ID NO: 1).  
           [0070]    Also provided by this aspect of the invention is an isolated WssH protein comprising the sequence of amino acids illustrated in FIG. 10 (SEQ ID NO: 9). The invention further provides a WssH protein encoded by a nucleic acid molecule according to the invention.  
           [0071]    The amino acid sequence shown in FIG. 10 (SEQ ID NO: 9) is a predicted translation of part of the longest open reading frame of the  Pseudomonas fluorescens  wssH gene. Translation of the WssH protein in vivo is expected to initiate at the first possible in-frame methionine codon (amino acid residue number 1 in the wssH translated sequence).  
           [0072]    The invention further provides an isolated nucleic acid molecule encoding a WssI protein, said protein comprising the sequence of amino acids illustrated in FIG. 11 (SEQ ID NO: 10). Preferably the nucleic acid molecule comprises the sequence of nucleotides from position 15751 to 16875 of the nucleic acid sequence illustrated in FIG. 2 (SEQ ID NO: 1).  
           [0073]    Also provided by this aspect of the invention is an isolated WssI protein comprising the sequence of amino acids illustrated in FIG. 11 (SEQ ID NO: 10). The invention further provides a WssI protein encoded by a nucleic acid molecule according to the invention.  
           [0074]    The amino acid sequence shown in FIG. 11 (SEQ ID NO: 10) is a predicted translation of part of the longest open reading frame of the  Pseudomonas fluorescens  wssI gene. Translation of the WssI protein in vivo is expected to initiate at the first possible in-frame methionine codon (amino acid residue number 1 in the wssI translated sequence).  
           [0075]    The invention further provides an isolated nucleic acid molecule encoding a WssJ protein, said protein comprising the sequence of amino acids from position 39 to position 324 of the amino acid sequence illustrated in FIG. 12 (SEQ ID NO: 11) or from position 2 to position 324 of the amino acid sequence illustrated in FIG. 12 (SEQ ID NO: 11). Preferably the nucleic acid molecule comprises the sequence of nucleotides from position 16938 to 17912 or from position 17052 to 17912 of the nucleic acid sequence illustrated in FIG. 2 (SEQ ID NO: 1).  
           [0076]    Also provided by this aspect of the invention is a WssJ protein comprising the sequence of amino acids from position 39 to position 324 of the amino acid sequence illustrated in FIG. 12 (SEQ ID NO: 11) or from position 2 to position 324 of the amino acid sequence illustrated in FIG. 12 (SEQ ID NO: 11). The invention further provides a WssJ protein encoded by a nucleic acid molecule according to the invention.  
           [0077]    The amino acid sequence shown in FIG. 12 (SEQ ID NO: 11) is the predicted translation of part of the longest open reading frame of the  Pseudomonas fluorescens  wssJ gene. Translation of the WssJ protein in vivo is expected to initiate at the first possible in-frame methionine codon (amino acid residue number 39 in the wssJ translated sequence). However, it is possible that translation may initiate at the first in-frame valine codon (amino acid residue number 1 in the wssJ translated sequence). In this case, the initiating amino acid of the protein product would still be methionine, not valine. Hence, the invention also provides a WssJ protein comprising the amino acid sequence from position 2 to position 324 of the amino acid sequence illustrated in FIG. 12 (SEQ ID NO: 11) but having an additional N-terminal methionine residue.  
           [0078]    As discussed below, the products of the wss G, H and I genes do not form part of the polysaccharide synthase complex but are thought to be involved in modification of the glucan-like polymer.  
           [0079]    In addition to wss operon, the inventors have identified a further novel operon in  P. fluorescens,  denoted the wsp operon, which encodes a chemotaxis-like operon of seven genes, wspA-F and wspR. The wspR gene product is involved in the regulation of the polysaccharide biosynthetic pathway of  P. fluorescens . The wspR gene product has also been found to be involved in bacterial attachment and biofilm development in  P. fluorescens . In addition, a wsp operon homologue has been shown to be involved in bacterial attachment in  P. aeruginosa  PA01.  
           [0080]    Isolated individual genes from the  Pseudomonas fluorescens  wsp operon are also encompassed within the scope of the invention, as are constructs comprising combinations of two or more of the individual genes. Thus, the invention provides an isolated nucleic acid molecule encoding a WspA protein, said protein comprising the sequence of amino acids illustrated in SEQ ID NO: 28. Preferably the nucleic acid molecule comprises the sequence of nucleotides from position 4535 to position 6178 of the nucleic acid sequence illustrated in SEQ ID NO: 27.  
           [0081]    Also provided by this aspect of the invention is a WspA protein comprising the sequence of amino acids illustrated SEQ ID NO: 28. The invention further provides a WspA protein encoded by a nucleic acid molecule according to the invention.  
           [0082]    The amino acid sequence shown in SEQ ID NO: 28 is the predicted translation of the longest open reading frame of the  Pseudomonas fluorescens  wspA gene. Translation of the WspA protein in vivo is expected to initiate at the first possible in-frame methionine codon.  
           [0083]    The invention also provides an isolated nucleic acid molecule encoding a WspB protein, said protein comprising the sequence of amino acids illustrated in SEQ ID NO: 29. Preferably the nucleic acid molecule comprises the sequence of nucleotides from position 6178 to position 6690 of the nucleic acid sequence illustrated in SEQ ID NO: 27.  
           [0084]    Also provided by this aspect of the invention is a WspA protein comprising the sequence of amino acids illustrated SEQ ID NO: 29. The invention further provides a WspB protein encoded by a nucleic acid molecule according to the invention.  
           [0085]    The amino acid sequence shown in SEQ ID NO: 29 is the predicted translation of the longest open reading frame of the  Pseudomonas fluorescens  wspB gene. Translation of the WspB protein in vivo is expected to initiate at the first possible in-frame methionine codon.  
           [0086]    The invention also provides an isolated nucleic acid molecule encoding a WspC protein, said protein comprising the sequence of amino acids illustrated in SEQ ID NO: 30. Preferably the nucleic acid molecule comprises the sequence of nucleotides from position 6687 to position 7946 of the nucleic acid sequence illustrated in SEQ ID NO: 27.  
           [0087]    Also provided by this aspect of the invention is a WspC protein comprising the sequence of amino acids illustrated SEQ ID NO: 30. The invention further provides a WspC protein encoded by a nucleic acid molecule according to the invention.  
           [0088]    The amino acid sequence shown in SEQ ID NO: 30 is the predicted translation of the longest open reading frame of the  Pseudomonas fluorescens  wspC gene. Translation of the WspC protein in vivo is expected to initiate at the first possible in-frame methionine codon.  
           [0089]    The invention provides an isolated nucleic acid molecule encoding a WspD protein, said protein comprising the sequence of amino acids illustrated in SEQ ID NO: 31. Preferably the nucleic acid molecule comprises the sequence of nucleotides from position 7943 to position 8641 of the nucleic acid sequence illustrated in SEQ ID NO: 27.  
           [0090]    Also provided by this aspect of the invention is a WspD protein comprising the sequence of amino acids illustrated SEQ ID NO: 31. The invention further provides a WspD protein encoded by a nucleic acid molecule according to the invention.  
           [0091]    The amino acid sequence shown in SEQ ID NO: 31 is the predicted translation of the longest open reading frame of the  Pseudomonas fluorescens  wspD gene. Translation of the WspD protein in vivo is expected to initiate at the first possible in-frame methionine codon.  
           [0092]    The invention further provides an isolated nucleic acid molecule encoding a WspE protein, said protein comprising the sequence of amino acids illustrated in SEQ ID NO: 32. Preferably the nucleic acid molecule comprises the sequence of nucleotides from position 8638 to position 10905 of the nucleic acid sequence illustrated in SEQ ID NO: 27.  
           [0093]    Also provided by this aspect of the invention is a WspE protein comprising the sequence of amino acids illustrated SEQ ID NO: 32. The invention further provides a WssA protein encoded by a nucleic acid molecule according to the invention.  
           [0094]    The amino acid sequence shown in SEQ ID NO: 32 is the predicted translation of the longest open reading frame of the  Pseudomonas fluorescens  wspE gene. Translation of the WspE protein in vivo is expected to initiate at the first possible in-frame methionine codon.  
           [0095]    The invention also provides an isolated nucleic acid molecule encoding a WspF protein, said protein comprising the sequence of amino acids illustrated in SEQ ID NO: 33. Preferably the nucleic acid molecule comprises the sequence of nucleotides from position 10902 to position 11912 of the nucleic acid sequence illustrated in SEQ ID NO: 27.  
           [0096]    Also provided by this aspect of the invention is a WspF protein comprising the sequence of amino acids illustrated SEQ ID NO: 33. The invention further provides a WspF protein encoded by a nucleic acid molecule according to the invention.  
           [0097]    The amino acid sequence shown in SEQ ID NO: 33 is the predicted translation of the longest open reading frame of the  Pseudomonas fluorescens  wspF gene. Translation of the WspF protein in vivo is expected to initiate at the first possible in-frame methionine codon.  
           [0098]    The invention still further provides an isolated nucleic acid molecule encoding a WspR protein, said protein comprising the sequence of amino acids illustrated in FIG. 13 (SEQ ID NO: 12). Preferably the nucleic acid molecule comprises the sequence of nucleotides illustrated in FIG. 14 (SEQ ID NO: 13).  
           [0099]    Also provided by this aspect of the invention is a WspR protein having the sequence of amino acids illustrated in FIG. 13 (SEQ ID NO: 12). The invention further provides a WspR protein encoded by a nucleic acid molecule according to the invention.  
           [0100]    The amino acid sequence shown in FIG. 13 (SEQ ID NO: 12) is the predicted translation of part of the longest open reading frame of the  Pseudomonas fluorescens  wspR gene. Translation of the WspR protein in vivo is predicted to initiate at the first possible in-frame methionine codon (amino acid residue number 1 in the wspR translated sequence).  
           [0101]    The wspR protein plays a role in the regulation of the wss operon and is essential for the production of the cellulose-like polysaccharide. Evidence for this comes from the glucan-like polysaccharide defective phenotype of wspR mutants. WspR has two domains (N and C) and a linker region. The N-terminus is highly similar to the N-terminus found in response regulator proteins. The C-terminus is widespread among prokaryotes, with many genomes containing a large number of genes with this C-terminus. However, the function of this domain is unknown. The only similar gene for which a phenotype has been assigned is PleD, from  Caulobacter cresentus  which is a cell-cycle gene and is essential for flagella ejection. PleD differs from WspR in that it has a duplicated N-terminal domain.  
           [0102]    The wild-type wspR gene (allele), the coding sequence of which is shown in FIG. 14 (SEQ ID NO: 13), is also referred to as the wspR-12 allele. Additional variants of wspR have been sequenced and are given different allele numbers, as illustrated in the accompanying Figures. Therefore, in addition to the wild-type wspR, the invention further provides allelic variants of wspR comprising the nucleotide sequences shown in FIGS.  16  (wspR-5; SEQ ID NO:15), 18 (wspR-9; SEQ ID NO:17), 20 (wspR-13; SEQ ID NO:19), 22 (wspR-14; SEQ ID NO:21) and 24 (wspR-19; SEQ ID NO:23). The sequences illustrated in these Figures are the coding regions only of the wspR alleles, from the predicted initiation codon to the first in-frame stop codon. The invention also provides isolated WspR proteins encoded by each of the variant wspR alleles, the amino acid sequences of these proteins being illustrated in FIGS. 15, 17,  19 ,  21  and  23  (SEQ ID Nos:14, 16, 18, 20 and 22, respectively).  
           [0103]    The invention also provides two isolated  P. fluorescens  genes which do not form part of the Wss operon but which may be involved in polysaccharide biosynthesis, specifically in chemical modification of glucan-like polymers, and also the protein products encoded thereby. These genes are designated mreB and pgi.  
           [0104]    Thus, the invention provides an isolated nucleic acid molecule encoding a  P. fluorescens  phosphoglucose isomerase protein, which nucleic acid molecule comprises the nucleotide sequence illustrated in FIG. 25 (SEQ ID NO:24).  
           [0105]    The invention further provides an isolated nucleic acid molecule comprising the nucleotide sequence illustrated in FIG. 26 (SEQ ID NO:25). The sequence shown in FIG. 26 (SEQ ID NO:25) covers a central region of the  P. fluorescens  mreB gene. A BLASTX search of genetic databases (via the BLAST server at www.ncbi.nlm.nih.gov/blast) identified the protein encoded from this region of DNA as being homologous to other sequenced mreB genes (e.g.  E. coli  mreB).  
           [0106]    The involvement of mre and pgi in polysaccharide expression in the  P. fluorescens  SBW25 LSWS mutant was initially determined through the isolation of mini-Tn5 transposon mutants (WS-12, pgi mutant; WS-39, mre mutant). Subsequent sequence analysis allowed the mapping of the mini-Tn5 insertion site into the genome, and the identification of the disrupted gene. The mreB mutant has a SM-like colony morphology, i.e. is never wrinkly, and binds the Congo Red stain on all media. The pgi mutant binds congo red on LB plates, but not on KB and is wrinkly on LB but not on KB. Furthermore, the pgi mutant was unable to form biofilm mats in microcosm vials, and the mat formed by the mre mutant was significantly weaker than that formed by the LSWS ancestor.  
           [0107]    The structure of the wss operon suggests that there are at least five genes products required for the expression of the glucan-like polysaccharide. Four of these genes (wssB-E) are required to form a polysaccharide synthase complex (based on homologies with the genes forming the cellulose synthase complex from  Acetobacter xylinus;  however,  A. xylinus  does not have a wssA homologue).  
           [0108]    Phosphoglucose isomerase (PGI, pgi) is a highly conserved enzyme found throughout the prokaryota and eukaryota. Its role is the interconversion of glucose-6-phosphate and fructose-6-phosphate. This interconversion is a critical step in the glucogenic and glucolytic pathways. With respect to the growth of  P. fluorescens  in media containing glycerol as the main carbon source, PGI is needed to transfer some of the energy flowing into the Embden-Meyerhof (EM) pathway from the utilisation of glycerol into the production of glucose, a necessary intermediate for polysaccharide biosynthesis.  
           [0109]    Because of the role of PGI in the production of intermediates for polysaccharide biosynthesis, the pgi gene may indirectly influence the efficiency and activity of the wss operon. It is thus a target for chemical mutation or other intervention as a means of indirectly influencing the activity of the wss operon.  
           [0110]    The invention further provides a candidate glucan-like polysaccharide producing operon isolated from  E. coli  as well as individual genes therefrom and the protein products encoded by these genes.  
           [0111]    The candidate  E. coli  polysaccharide producing operon provided by the invention comprises the nucleotide sequence illustrated in FIG. 27 (SEQ ID NO:26). The co-ordinates for the individual genes within the DNA fragment shown in FIG. 2 (SEQ ID NO:1) are as follows: yhjQ 336-1070 (wssA homologue); yhjO 995-3685 (wssB homologue); yhjN 3591-6035 (wssC homologue); yhjM 6036-7148 (wssD homologue); yhjL 7051-10602 (wssE homologue); yhjK 10627-12672; and yctA 12819-past the end of the given sequence.  
           [0112]    Examination of the  E. coli  genome (via the public NCBI  E. coli  database, see: www.ncbi.nlm.nih.gov/cgi-bin/Entrez/framik?db=Genome&amp;gi=115); identified an operon (yhj) which had five genes with strong homology to the wssA-E genes of the  P. fluorescens  polysaccharide biosynthetic operon. The yhj operon (yhjK-Q) is located at the 3,694,861-3,679,861 region of the  E. coli  genome. This region also includes dctA. At the time the inventors identified this operon there had been no previous publication reporting the expression of cellulose or a modified cellulose from this operon in  E. coli . Nevertheless, by virtue of the fact that  E. coli  appeared to have the necessary genes for cellulose expression, the inventors decided to test whether a polysaccharide-producing mutant strain could be generated using the microcosm system used to generate a  P. fluorescens  strain that expressed a glucan-like polysaccharide. Subsequently, a WS-like  E. coli  DH5A mutant was isolated which takes up Congo Red stain on agar plates, and which binds the cellulose specific Calcoflour stain. These initial tests strongly support the suggestion that this strain of  E. coli  now expresses polysaccharide in a similar manner to the LSWS strain of  P. fluorescens  SBW25. Zogaj et al. (Zogaj, X., Nimtz, M., Rohde, M., Bokranz, W., and Romling, U. (2001). The multicellular morphotypes of  Salmonella typhimurium  and  Escherichia coli  produce cellulose as the second component of the extracellular matrix. Mol. Microbiol. 39: 1452-1463) have subsequently also described the expression of cellulose in  E. coli.    
           [0113]    In accordance with the present invention, a defined nucleic acid includes not only the identical nucleic acid but also any minor base variations, including in particular base substitutions which result in a synonymous codon (a different codon specifying the same amino acid residue) due to the degeneracy of the genetic code. The term “nucleic acid” includes single or double stranded RNA, single or double stranded DNA, synthetic forms and mixed polymers, both sense and antisense strands, and may be chemically or biochemically modified or may contain non-natural or derivatized nucleotide bases as will be readily appreciated by those skilled in the art. Such modifications include, for example, labels, methylation, substitution of one or more naturally occurring nucleotide bases with an analog. Libraries of bacterial chromosomal fragments may be screened as natural sources of the nucleic acids of the present invention. Alternatively, nucleic acid sequences according to the invention may be produced using recombinant or synthetic means, for example by PCR amplification of sequences resident in chromosomal DNA or cloned fragments thereof. Generally such techniques are well known in the art (see Sambrook et al. (1989),  Molecular Cloning: A Laboratory Manual,  Cold Spring Harbor Laboratory Press; F. M. Ausubel et al. (eds.),  Current Protocols in Molecular Biology,  John Wiley &amp; Sons, Inc. (1994)).  
           [0114]    The DNA molecules according to the invention may, advantageously, be included in a suitable expression vector to express the protein encoded therefrom in a suitable host. Procedures for incorporation of a cloned DNA into a suitable expression vector, transformation of a host cell and subsequent selection of the transformed cells are well known to those skilled in the art, as provided by Sambrook et al. (1989),  Molecular Cloning: A Laboratory Manual,  Cold Spring Harbor Laboratory Press or F. M. Ausubel et al. (eds.),  Current Protocols in Molecular Biology,  John Wiley &amp; Sons, Inc. (1994).  
           [0115]    An expression vector according to the invention includes a vector comprising a nucleic acid molecule according to the invention operably linked to regulatory sequences, such as promoter regions, that are capable of effecting expression of the said nucleic acid. The term “operably linked” refers to a juxtaposition wherein the components described are in a relationship permitting them to function in their intended manner. Such vectors may be introduced into a suitable host cell to provide for expression of a polypeptide according to the invention. Thus, in a further aspect, the invention provides a process for preparing polypeptides according to the invention which comprises cultivating a host cell, comprising an expression vector as described above under conditions to provide for expression by the vector of a coding sequence encoding the polypeptides, and recovering the expressed polypeptides.  
           [0116]    Expression vectors suitable for driving expression of a given protein in a prokaryotic host cell may be, for example, plasmid or phage vectors provided with an origin of replication, and a promoter to drive transcription of mRNA encoding the said protein and optionally a regulator of the promoter. The vectors may contain one or more selectable markers, such as, for example, ampicillin resistance.  
           [0117]    Sequence elements required for prokaryotic expression include promoter sequences to bind RNA polymerase and processing information sites such as ribosome binding sites, transcription termination sites etc. For example, a bacterial expression vector may include a promoter to bind RNA polymerase and direct an appropriate frequency of transcription initiation at the transcription start site and for translation initiation the Shine-Dalgarno sequence and a translation initiation codon (usually AUG). The promoter may be the promoter naturally associated with the coding region in question or may be a heterologous promoter such as, for example, the lac promoter. In a preferred embodiment the promoter is inducible, being activated by binding of an appropriate transcriptional regulatory molecule or promoter-specific RNA polymerase. Expression of the target protein can thus be temporally controlled by controlling expression and/or activation of the transcriptional regulatory molecule or promoter-specific RNA polymerase. Well known examples of such inducible systems are the T7 promoter/T7polymerase, T3 promoter/T3 polymerase and SP6 promoter/SP6 polymerase systems and the lacUV5 promoter/IPTG system. Vectors suitable for expression in a range of prokaryotic hosts may be obtained commercially or assembled from the sequences described by methods well known in the art.  
           [0118]    A further aspect the invention also provides a host cell or organism comprising an expression vector according to the invention. Preferably, the host cell/organism is a prokaryotic cell/organism.  
           [0119]    In accordance with the present invention, a defined protein or polypeptide includes proteins which are substantially homologous but have one or more conservative amino acid changes, including naturally occurring allelic variants, or in vivo or in vitro chemical or biochemical modifications (e.g. acetylation, carboxylation, phosphorylation, glycosylation etc). In this context, a “substantially homologous” sequence is regarded as a sequence which shares at least 80%, preferably at least 90% and more preferably at least 95% amino acid sequence identity with the proteins or polypeptides of the invention. The protein according to the invention may be recombinant, synthetic or naturally occurring, but is preferably recombinant.  
           [0120]    Also within the scope of the invention are fusion proteins/polypeptides comprising a protein according to the invention. The proteins of the invention may be fused either N-terminally or C-terminally to heterologous protein or peptide fragments, for example to facilitate purification of the fusion protein. Fusion proteins will typically be made by recombinant nucleic acid techniques or may be chemically synthesized.  
           [0121]    The invention also provides for cleavage fragments of the full length proteins, particularly cleavage fragments of the enzymes which share homology with cellulose synthase subunits. It is not uncommon for bacterial genes to encode a inactive precursor form of an enzyme/enzyme subunit which is post-translationally processed to yield shorter polypeptides which participate in catalytic and/or regulatory activity of the enzyme/enzyme complex.  
           [0122]    In a still further aspect, the invention provides a method of constructing an exopolysaccharide-producing bacterial strain, which method comprises introducing an expression vector suitable for overexpression of a WspR protein into a host bacterial strain, the genome of which contains a wss-like operon.  
           [0123]    Advantageously, the host bacterial strain may be a Pseudomonas strain or an  E. coli  strain. Preferred host strains include wild type Pseudomonas fluorescens strain SBW25 and  E. coli  strain K12.  
           [0124]    In one embodiment, the WspR protein is a wild-type WspR protein comprising the amino acid sequence illustrated in FIG. 13 (SEQ ID NO:12), in which case optimum production of polysaccharide from the resultant exopolysaccharide-producing strain may require the addition of NaCl to the culture medium (see accompanying Examples). In this embodiment, the expression vector may conveniently comprise the sequence of nucleotides shown in FIG. 14 (SEQ ID NO:13) operably linked to sequences which control its expression.  
           [0125]    In a further embodiment, the WspR protein is an allelic variant of the  Pseudomonas fluorescens  WspR protein having the amino acid sequence illustrated in FIG. 21 (WspR-14; SEQ ID NO:20) or in FIG. 23 (WspR-19; SEQ ID NO:22). Exopolysaccharide-producing strains expressing these variant WspR proteins generally do not require the presence of additional NaCl for optimal polysaccharide production (see accompanying Examples).  
           [0126]    For this aspect of the invention, it may be useful to construct an expression vector in which nucleic acid encoding the wspR protein is placed under the control of an inducible promoter (see above). Host cells containing such a construct can be grown up in culture with wspR expression, and hence β-linked glucan production, switched off then at the appropriate time expression of wspR can be induced, leading to expression of the cellulose biosynthetic enzymes.  
           [0127]    Also included within the scope of the invention is an embodiment in which the expression vector is an invasive plasmid which can be used to transform a host bacterium in situ, meaning in the field or in the natural environment of the bacterium as opposed to in in vitro culture in a laboratory.  
           [0128]    A particularly useful application of this method of the invention is in the introduction of a WspR expressing plasmid into a soil-dwelling bacterium. Switching on glucan-like polysaccharide production in such a bacterium may enable the bacterium to stick to and colonise the roots of a plant or may render the bacterium more resistant to dessication. Cellulose production is known in a number of bacteria, but has received attention only in Acetobacter (where it has been studied from the point of view of cellulose extraction) and Agrobacterium. Studies in Agrobacterium have focussed on its role in attachment to the plant surface. The data is not entirely clear, but it does appear to have a role. The inventors postulate that the ecological role of the glucan-like polysaccharide in  P. fluorescens  may be in surface colonisation rather than attachment. It is proposed that the wild-type bacterium can regulate (through the wsp (chemosensory) operon) the amount of cellulose produced from the poles of the cells and can thereby control whether cells spread rapidly across a surface in a thin later, or pile up in a mound. The ability to control surface colonisation may have considerable biotechnological potential.  
           [0129]    The invention further provides an exopolysaccharide-producing strain which is obtainable by the above-described methods. In a preferred embodiment, the exopolysaccharide-producing strain, in addition to expression of a WspR protein, further carries a mutation in the mreB gene, the pgi gene or both the aforementioned genes.  
           [0130]    As explained herein, both the mreB gene product and the pgi gene product of  Pseudomonas fluorescens , and hence homologous genes from other bacterial strains, may influence the nature of the polysaccharide product produced by the action of enzymes encoded by the wss operon. Hence, strains which carry a mutation in either or both of these genes may produce varying polysaccharide products.  
           [0131]    The wspR protein also plays a role in the attachment of bacteria to the sides and surfaces of culture containers. Bacterial attachment is the first stage in the development of a biofilm. Subsequent biofilm development proceeds from the attached cells out into the liquid media and new bacterial remain connected to the attached cells via the expression of exocellular polysaccharide or proteinaceous matrix or skeleton. Evidence for this in  P. fluorescens  comes from a comparison of attachment abilities of various wrinkley spreader mutants. The original wrinkly spreader strain (WS) and the mutant strain WS-13 (unable to express glucan-like polysaccharide) are able to attach readily to the surfaces of culture containers. In contrast, WS-4 (wspR − ) is unable to attach to surfaces at all. Similarily, a wrinkly spreader strain with a wspΔ (WS wspΔ) is not able to attach either. Futhermore, like  P. fluorsecens  WS wspΔ, the inventors have found that  P. aeruginosa  PA01 deleted for the wsp-like operon, is also defficient in bacterial attachment.  
           [0132]    A particularly useful application of the invention may be found in the removal of a biofilm. The production of extracellular cellulose plays an important part in the development of biofilms. The inventors have observed that expression of the WspR allelic variants WspR-5, WspR-9 and WspR-13 in a strain of  Pseudomonas fluorescens  which is producing glucan-like polysaccharide (i e having wrinkly spreader morphology) results in cessation of polysaccharide production and return to an SM phenotype. It is therefore to be expected that introduction of an expression vector encoding wspR-5, wspR-9 or wspR-13 into bacteria which make up a biofilm would shut down polysaccharide production and hence promote disruption of the biofilm. Again, this would be advantageously achieved using an invasive plasmid encoding wspR-5, wspR-9 or wspR-13 to transform the biofilm in situ. This approach might be adapted for use in essentially any situation where formation of biofilms is known to be a problem. A particular example from industry is in the paper making process where a number of rolling and screening stages are used, each stage being subject to bio-fouling, often associated with formation of a biofilm. Currently this problem is addressed with the use of anti-microbial agents.  
           [0133]    In addition, the identification of the role of WspR in bacterial attachment and biofilm development provides a further application of appropriately modified  P. fluorescens  or  P. aeruginosa  strains in which bacetrial attachment is used to screen chemical and pharmaceutical libraries for compounds that inhibit attachment and biofilm growth. These compounds might directly prevent cellulose production, directly prevent normal WspR function, or prevent WspR function indirectly. The compounds may bind specifically with WspR preventing normal wspR interactions with other cellular components involved in cellulose biosythesis, attachment or biofilm development. Alternatively, these compounds may interfere with the production or function of cellular components which act on, or with WspR for normal function. The screening system can also be used to identify compounds inhibiting cellulose biosythesis, attachment or biofilm development without interacting directly with WspR.  
           [0134]    Screening Assays for Inhibitors of Exopolysaccharide Production, Bacterial Attachment and Biofilm Development.  
           [0135]    Using  P. fluorescens  SBW25 and  P. aeruginosa  PA01, the present inventors have discovered that the wsp operon plays an important role in the attachment of bacterial cells to solid surfaces. Further, it has been determined that the development of biofilms first requires bacterial attachment. Biofilms are of particular problem in human medicine, where high concentrations of bacteria can exist in tissues and be protected by a secreted, biofilm matrix or scaffold. The inventors&#39; observations lead to the conclusion that in  P. fluorescens, P. aeruginosa  and other pseudomonads bacterial attachment is regulated by wsp, or a wsp-like operon of speacilised chemotatic or cheomosensory genes. Some environmental or internal signal is received by this system, and the signal is passed down to WspR, or a WspR-like protein. The activated WspR then stimulates the expression of downstream genes required for the actual physical attachment of bacteria.  
           [0136]    The observation that the wsp operon is involved in bacterial attachment and biofilm development has led to the development of assays which may be used to screen libraries of compounds to identify inhibitors of expolysaccharide production, bacterial attachment and biofilm development. In essence, these assays rely on the production of exopolysaccharide by wrinkly spreader (WS) strains or modified SM strains.  
           [0137]    In  P. fluorescens , attachment and activated WspR, also stimulate the expression of exopolysaccharide leading to the development of a biofilm. In  P. aeruginosa  PA01, no cellulose biosynthetic genes exist but it is possible that activated WspR may induce other biofilm-forming genes, such as alignate biosythesis.  
           [0138]    A number of assays are provided which may be used to screen chemical libraries for compounds that inhibit bacterial attachment and biofilm development:  
           [0139]    Assay for Exopolysaccharide Production:  
           [0140]    In this assay exopolysaccharide-producing bacteria are incubated in the presence of test chemicals on agar plates containing a dye which specifically stains the exopolysaccharide, for example Congo Red. Production of the exopolysaccharide is scored by observing uptake of the dye.  
           [0141]    Assay for attachment and biofilm Production:  
           [0142]    In this assay exopolysaccharide-producing bacteria are incubated in the presence of test chemicals in liquid broth culture. Attachment and biofilm development is scored by visual inspection, as described in the accompanying examples. Crystal violet staining may also be used to provide quantitive results for bacterial attachment.  
           [0143]    Advantageously, the attachment assay may be carried out in microtitre assay plates where 96 or more individual cultures can be tested on a single plate. The testing of different culture containers is important in assay optimisation, as some attachment systems are affected by different materials ( P. fluorescens  SBW25 will attach easily to glass and polystyrene).  
           [0144]    The exopolysaccharide production assay and the assay for attachment and biofilm formation may both be performed using any of the exopolysaccharide producing bacterial strains described herein, including evolved variant strains having wrinkly spreader morphology, recombinant strains, mutagenized strains etc.  
           [0145]    The assays can also be perfomed using modified forms of the wild-type  P. fluorescens  SBW25 which have been engineered to express either wpR14 or wspR19. The wild-type SBW25 strain contains the wild-type wspR12 allele and is phenotypically smooth (SM). When modified to express wspR14 or wspR19 it exhibits the wrinkly spreader phenotype, produces exopolysaccharide and forms biofilms. In contrast, if a wrinkly spreader strain is engineered to wspR5, 9 or 13 it will exhibit a smooth (SM) phenotype. These alterations can be used to manipulate the assay an enable screening on specific allelic forms of wspR, for example a wrinkly spreader strain engineered to express wspR5 (now phenotypically SM) will produce exopolysaccharide if the test chemical interferes with the wspR5 protein but not the chromosomal copy of wspR12.  
           [0146]    A liquid culture based assay for inhibitors of attachment may also be carried out using a bacterial strain which expresses the gene-products of a wsp-like operon and a control strain which is essentially identical but which does not expresses the gene-products of the wsp-like operon. The inventors have shown by experiment that Wsp homologues are required for attachment in the liquid culture system. Thus, the assay can be carried out using essentially any bacterial strain which expresses the Wsp homologs required for attachment. Specificity is provided by the use of the control strain which does not express the Wsp homologues. In this context the term “Wsp homologues” encompasses proteins which exhibit at least 75% sequence similarlity with the homologous  P. fluorescens  Wsp protein and/or the homologous  P. aeruginosa  Wsp protein at the amino acid level.  
           [0147]    “Wsp-like operon” is a generic term used herein to describe a novel class of bacterial operons containing genes which encode proteins involved in the regulation of glucan-like polysaccharides synthesis, in bacterial attachment and/or biofilm formation. Encompassed within the term “wsp-like operon” are operons which are homologous to the wsp operon of  Pseudomonas fluorescens , the complete nucleotide sequence of which is given herein. Preferably, the wsp-like operon comprises a sequence of nucleotides which shares at least 50% nucleotide sequence identity with the sequence of nucleotides shown in SEQ ID NO: 27. Even more preferably, the wss-like operon may comprise a sequence of nucleotides at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 90% or at least 95% identical to the  P. fluorescens  wsp operon shown in SEQ ID NO: 27. Percentage nucleotide identity may be calculated by comparing entire wsp operon sequences or by comparing the coding regions of the individual genes of the operons. In the latter case, the coding regions of homologous genes should be compared. A value for the overall percentage sequence identity may then be derived by taking an average over all the individual homologous coding regions. A complete annotation of the  P. fluorescens  wsp operon, showing the positions of the coding regions is listed elsewhere in this specification.  
           [0148]    In a preferred embodiment the assay may be based on the use of  P. aeruginosa  strain PA01 (a well-characterised strain which is available from public strain collections, see Holloway, B. W. (1955). Genetic recombination in  Pseudomonas aeruginosa. J. Gen. Microbiol.  13, 572-581; Stover, C. K., Pham, X. Q., Erwin, A. L., et al. (2000). Complete genome sequence of  Pseudomonas aeruginosa  PA01, an opportunistic pathogen. Nature 406: 959-964) in order to screen for chemicals which specifically interfere with PA01 attachment. By using wild type PA01 and PA01 wspΔ (i.e. PA01 with the wsp operon deleted) it is possibly to identify chemicals which specifically interfere with attachment of  P. aeruginosa . Experiments have shown that the PA01 wsp operon homologue is required for attachment in the liquid culture system, thus PA01 wspΔ should always grow in liquid culture unless the test chemicals are toxic but does not attach, whereas wild type PA01 should attach unless the test chemical has an effect on the attachment process.  
           [0149]    In a further preferred embodiment the attachment assay may be carried out using wild-type  P. fluorescens  and an equivalent strain deleted for the wsp operon, e.g. wild-type  P. fluorescens  SBW25 and  P. fluorescens  SBW25 wspΔ.  
           [0150]    A wide variety of candidate chemicals may be tested in the screening methods of the invention. Suitable test chemicals may include, for example, chemicals having a known biochemical activity, chemicals having no such identified activity and completely new molecules or libraries of molecules such as might be generated by combinatorial chemistry. Test chemicals which are nucleic acids, including naturally occuring nucleic acids and synthetic analogues, polypeptides or proteins are not excluded.  
           [0151]    Typically, screening assays involve running a plurality of assay mixtures in parallel with different concentrations of the test chemical. Typically, one of these concentrations serves as a negative control, i.e. zero concentration of test chemical.  
           [0152]    Construction of Exopolysaccharide-Producing Strains Based on Expression of wss Gene Products  
           [0153]    In a further aspect, the invention provides exopolysaccharide-producing bacterial strains based on expression of the wss operon.  
           [0154]    It will be readily appreciated by a skilled artisan that it is possible to genetically modify a bacterium which does not naturally produce extracellular polysaccharide to make an exopolysaccharide-producing strain by introducing an expression vector suitable for driving expression of protein products of the wss operon which are required for polysaccharide biosynthesis.  
           [0155]    Accordingly, the invention provides an exopolysaccharide-producing bacterial strain which is a bacterial host strain containing an expression vector including a nucleic acid comprising coding regions of a wss-like operon operably linked to regulatory sequences which control expression of the said nucleic acid.  
           [0156]    In one embodiment, the expression vector may comprise a nucleic acid comprising all the coding regions of the  Pseudomonas fluorescens  wss operon, or all the coding regions of the  E. coli  yhj operon operably linked to appropriate expression regulatory sequences, or just the coding regions which are absolutely essential for polysaccharide biosynthesis. It will be appreciated that the expression vector may also contain intergenic regions from the wss operon in question in addition to the coding regions. It will further be appreciated that exopolysaccharide producing strains could be produced by co-expressing wss gene products from different species/strains. By combining cellulase synthase subunits from different species/strains in this manner it may be possible to alter the specificity of the enzyme and hence alter the structure of the resultant polysaccharide.  
           [0157]    The expression regulatory sequences may comprise a promoter which is constitutively active in the bacterial host cell in question, leading to constitutive expression of the wss proteins or, in an alternative embodiment, an inducible promoter to enable polysaccharide production to be regulated.  
           [0158]    In one embodiment, the expression regulatory sequences comprise the “authentic” promoter region of the wss operon. For example, an expression vector containing coding regions from the  P. fluorescens  wss operon would contain the promoter region of the  P. fluorescens  wss operon. In one embodiment, the expression vector might comprise a substantially complete wss operon, including the promoter region and all of the enzyme-encoding genes.  
           [0159]    For those embodiments wherein the expression vector comprises an authentic wss promoter, the host bacterial strain should also comprise nucleic acid encoding a WspR protein which functions as a regulator of the wss promoter. Advantageously, the nucleic acid encoding the WspR protein may be present on a second expression vector under the control of a constitutive or inducible promoter, as appropriate.  
           [0160]    The wss operon contains several more genes than would, on the basis of homology with cellulose biosynthetic operons from other bacterial species, seem to be required for the production of ‘pure’ cellulose. Three genes at the end of the operon (wssG, wssH, wssI) show similarity to the  P. aeruginosa  genes, aglF, algI &amp; algJ. In  P. aeruginosa , these three genes are responsible for acetylation of mannose residues that are part of the alginate polymer. The similarity between these genes and those in  P. fluorescens  is low, nonetheless, their presence suggests that a basic glucan-like polymer may be modified, possibly by acetylation. Mutants in wssGHI have been isolated that have a different phenotype on agar plates (no longer wrinkly) and lack ability to colonise the air-liquid interface in static microcosms, and yet still produce a polysaccharide that stains positively with calcofluor and Congo red. Thus, there exist within the wss operon genes that affect the nature of the basic glucan-like polymer. This opens the door to the construction of novel glucan-like polysaccharides. Moreover, further genes which influence the nature of the glucan-like polymer may be found outside of the wss operon, a particular example being the pgi gene discussed previously. It may thus be possible to construct further novel glucan-like polysaccharides by manipulation of genes outside the wss operon, by mutation or other means.  
           [0161]    Process for the Production of Glucan-Like Polysaccharide  
           [0162]    In a further aspect the invention also provides a process for the production of glucan-like polysaccharide from an exopolysaccharide-producing bacterial strain, which process comprises the steps of growing an exopolysaccharide-producing bacterial strain according to the invention and isolating the polysaccharide produced thereby.  
           [0163]    The exopolysaccharide-producing bacterial strain can be any such strain described herein, including evolved variant strains, engineered strains, mutant strains etc.  
           [0164]    In a preferred embodiment, the step of isolating the polysaccharide comprises lysing the bacteria in a bacterial lysis solution, for example 20 mM Tris HCl pH 8.0, 5 mM MgCl 2 , 0.5% Sarkosyl, 1 mg/ml fresh lysozyme and incubating the sample thus obtained in with a second lysis solution comprising detergent and Proteinase K, for example 500 mM EDTA pH 9.0, 1% Sarkosyl, 1.5 mg/ml Proteinase K.  
           [0165]    As discussed above, exopolysaccharide-producing evolved variants of Pseudomonas, for example evolved variants of  Pseudomonas fluorescens , have a characteristic wrinkly spreader morphology, as described by Rainey &amp; Travisano, 1998, Nature, 394: 69-72. Cells of the WS morph adhere firmly to each other and to surfaces, allowing the formation of a self-supporting mat at the air-broth interface when grown in a standard microcosm (see Example 1). The WS morph can also be grown on hard agar plates to form single colonies. Polysaccharide can be isolated from cells grown in either type of culture, colonies or mats, using substantially the same protocol for polysaccharide purification (see Example 2).  
           [0166]    It will be appreciated that the precise composition of the polysaccharide product produced by a given exopolysaccharide-producing strain may vary slightly according to the type of carbohydrate added to the culture media in which the bacteria are grown, as this will ultimately determine the nature of the substrate available for the polysaccharide biosynthetic enzymes. Accordingly, it is within the scope of the invention to vary the precise composition of the polysaccharide by manipulating the type of carbohydrate added to the culture medium. 
       
    
    
       [0167]    The present invention will be further understood with reference to the following non-limiting Examples, together with the accompanying Figures in which:  
         [0168]    [0168]FIG. 1 is a schematic representation of the wss operon. The operon consists of ten genes (wssA-J) located on a ˜20 kb fragment of  Pseudomonas fluorescens  SBW25 genomic DNA. Some restriction sites are indicated below the coding regions; B, BamHI; H, HindIII and K, KpnI. The scale is given in 1 kb units.  
         [0169]    [0169]FIG. 2 shows the nucleotide sequence of a contiguous 20,306 bp fragment of genomic DNA from  Pseudomonas fluorescens  SBW25. The wss operon, encoding the cellulose biosynthetic genes and associated genes, is located approximately between 2,200-18,000 bp.  
         [0170]    [0170]FIG. 3 illustrates the predicted translation of the longest open reading frame of the  P. fluorescens  wssA gene. The sequence shown is from the first potential start codon (GTG; V valine) to the first in-frame stop codon. In order to easily place the peptide sequence within the DNA sequence of the wss operon the potential GTG-encoded start codon has been left as ‘V’ in the peptide sequence even though the initiating amino acid would in fact be methionine. The first potential ATG start codon (M) is also marked.  
         [0171]    [0171]FIG. 4 illustrates the predicted translation of the longest open reading frame of the P. fluorescens wssB gene, from the first potential ATG start codon to the first in-frame stop codon.  
         [0172]    [0172]FIG. 5 illustrates the predicted translation of the longest open reading frame of the  P. fluorescens  wssC gene, from the first potential GTG start codon (V) to the first in-frame stop codon. The first potential ATG start codon is also marked (M).  
         [0173]    [0173]FIG. 6 illustrates the predicted translation of the longest open reading frame of the  P. fluorescens  wssD gene, from the first potential GTG start codon (V) to the first in-frame stop codon. The first potential ATG start codon is also marked (M).  
         [0174]    [0174]FIG. 7 illustrates the predicted translation of the longest open reading frame of the  P. fluorescens  wssE gene, from the first potential ATG start codon (M) to the first in-frame stop codon.  
         [0175]    [0175]FIG. 8 illustrates the predicted translation of the longest open reading frame of the  P. fluorescens  wssF gene, from the first potential ATG start codon (M) to the first in-frame stop codon.  
         [0176]    [0176]FIG. 9 illustrates the predicted translation of the longest open reading frame of the  P. fluorescens  wssG gene, from the first potential ATG start codon (M) to the first in-frame stop codon.  
         [0177]    [0177]FIG. 10 illustrates the predicted translation of the longest open reading frame of the  P. fluorescens  wssH gene, from the first potential ATG start codon (M) to the first in-frame stop codon.  
         [0178]    [0178]FIG. 11 illustrates the predicted translation of the longest open reading frame of the  P. fluorescens  wssI gene, from the first potential ATG start codon (M) to the first in-frame stop codon.  
         [0179]    [0179]FIG. 12 illustrates the predicted translation of the longest open reading frame of the  P. fluorescens  wssJ gene, from the first potential ATG start codon (M) to the first in-frame stop codon.  
         [0180]    [0180]FIG. 13 illustrates the predicted translation of the longest open reading frame of the wild-type  P. fluorescens  wspR gene (allele WspR-12).  
         [0181]    [0181]FIG. 14 shows the nucleotide sequence of the coding region of the wild-type  P. fluorescens  wspR gene (allele WspR-12) from the first potential ATG start codon to the first in-frame stop codon TAG.  
         [0182]    [0182]FIG. 15 illustrates the predicted translation of the longest open reading frame of the variant  P. fluorescens  wspR allele WspR-5.  
         [0183]    [0183]FIG. 16 shows the nucleotide sequence of the coding region of the variant  P. fluorescens  wspr allele WspR-5 from the first potential ATG start codon to the first in-frame stop codon TAG.  
         [0184]    [0184]FIG. 17 illustrates the predicted translation of the longest open reading frame of the variant  P. fluorescens  wspR allele WspR-9.  
         [0185]    [0185]FIG. 18 shows the nucleotide sequence of the coding region of the variant  P. fluorescens  wspR allele WspR-9 from the first potential ATG start codon to the first in-frame stop codon TAG.  
         [0186]    [0186]FIG. 19 illustrates the predicted translation of the longest open reading frame of the variant  P. fluorescens  wspR allele WspR-13.  
         [0187]    [0187]FIG. 20 shows the nucleotide sequence of the coding region of the variant  P. fluorescens  wspr allele WspR-13 from the first potential ATG start codon to the first in-frame stop codon TAG.  
         [0188]    [0188]FIG. 21 illustrates the predicted translation of the longest open reading frame of the variant  P. fluorescens  wspR allele WspR-14.  
         [0189]    [0189]FIG. 22 shows the nucleotide sequence of the coding region of the variant  P. fluorescens  wspR allele WspR-14 from the first potential ATG start codon to the first in-frame stop codon TAG.  
         [0190]    [0190]FIG. 23 illustrates the predicted translation of the longest open reading frame of the variant  P. fluorescens  wspR allele WspR-19.  
         [0191]    [0191]FIG. 24 shows the nucleotide sequence of the coding region of the variant  P. fluorescens  wspR allele WspR-19 from the first potential ATG start codon to the first in-frame stop codon TAG.  
         [0192]    [0192]FIG. 25 shows the nucleotide sequence of a near-contiguous piece of DNA of 1,136 bp from the genome of Pseudomonas fluorescens SBW25. The sequence covers a central region of the pgi (phosphoglucose isomerase) gene. A BLASTX search of genetic databases (via the BLAST server at www ncbi nlm nih gov/blast) identified the protein encoded from this region of DNA as being homologous to other sequenced pgi genes (e.g.  E. coli  pgi).  
         [0193]    [0193]FIG. 26 shows the nucleotide sequence of a near-contiguous piece of DNA of 703 bp from the genome of  Pseudomonas fluorescens  SBW25. The sequence covers a central region of the mreB (murien biosynthesis B) gene. A BLASTX search of genetic databases (via the BLAST server at www ncbi nlm nih gov/blast) identified the protein encoded from this region of DNA as being homologous to other sequenced mreb genes (e.g.  E. coli  mreB).  
         [0194]    [0194]FIG. 27 shows the nucleotide sequence for a contiguous piece of DNA of 14,000 bp from the genome of  Escherichia coli . The DNA sequence described here has been obtained from the public NCBI  E. coli  database (see: www ncbi.nlm.nih.gov/cgi-bin/Entrez/framik?db=Genome&amp;gi=115); the 14,000 bp comes from the 3,694,861-3,679,861 region of genome. This region includes the yhj operon (yhjK-Q) and also includes dctA. The co-ordinates for the coding regions of the yhj genes can be found at the web site given above; the co-ordinates for the genes for the DNA segment shown in this Figure are: (any codon start)  
         [0195]    yhjQ 336-1070 (wssA homologue);  
         [0196]    yhjO 995-3685 (wssB homologue);  
         [0197]    yhjN 3591-6035 (wssC homologue);  
         [0198]    yhjM 6036-7148 (wssD homologue);  
         [0199]    yhjL 7051-10602 (wssE homologue);  
         [0200]    yhjK 10627-12672; and  
         [0201]    yctA 12819-past the end of the given sequence.  
         [0202]    [0202]FIG. 28 is a shematic representation of the  Pseudomonas fluorescencs  wsp operon. The operon consists of seven genes (wspA-F and wspR) located on a ˜10 kb fragment of  Pseudomonas fluorescens  SBW25 genomic DNA.  
         [0203]    [0203]FIG. 29 shows the nucleotide sequence of a fragment of chromosomal DNA from  Pseudomonas fluorescens  SBW25 including the wsp operon.  
         [0204]    A complete annotation for this sequence is given in the accompanying Examples.  
         [0205]    [0205]FIG. 30 illustrates the predicted translation of the longest open reading frame of the  P. fluorescens  wspA gene.  
         [0206]    [0206]FIG. 31 illustrates the predicted translation of the longest open reading frame of the  P. fluorescens  wspB gene.  
         [0207]    [0207]FIG. 32 illustrates the predicted translation of the longest open reading frame of the  P. fluorescens  wspC gene.  
         [0208]    [0208]FIG. 33 illustrates the predicted translation of the longest open reading frame of the  P. fluorescens  wspD gene.  
         [0209]    [0209]FIG. 34 illustrates the predicted translation of the longest open reading frame of the  P. fluorescens  wspE gene.  
         [0210]    [0210]FIG. 35 illustrates the predicted translation of the longest open reading frame of the  P. fluorescens  wspF gene.  
         [0211]    [0211]FIG. 36 illustrates the predicted translation of the longest open reading frame of the  P. fluorescens  wspR gene. 
     
    
       [0212]    Strain Nomenclature  
         [0213]    Throughout the present application various abbreviations are used for  P. fluorescens  SBW25 strains. The wild-type strain is sometimes referred to as “SM” because of its smooth colony morphology. The Wrinkly Spreader strain expresses glucan-like polysaccharide (GLP) and is often referred to as “WS” because of its wrinkled colony morphology. A variety of WS derivatives are also used and in general they are deficient in GLP production. They are specifically referred to when necessary using numbers, e.g. WS-4, WS-13 etc. In some cases, the same genetic mutation is present in both SM and WS genotypes. In this case, SM genotypes are referred to using the same numbering or naming system as for the original WS mutants (e.g. SM-13), or SM or WS are added to the genotype (e.g. SM wspΔ or WS wspΔ).  
       Example 1  
       [0214]    Obtaining Glucan-Like Polysaccharide Over-Producing  P. fluorescens.    
         [0215]    The following example uses the wild type  P. fluorescens  strain SBW25 which may be isolated from sugar beet leaves, as described by Rainey, P. B. &amp; Bailey, M. J., 1996 , Mol. Microbiol.,  19: 521-533, and propagated in King&#39;s medium B (KB).  P. fluorescens  SBW25 is also freely available from the culture collection at the Department of Plant Sciences, University of Oxford, Oxford, UK.  
         [0216]    In order to obtain polysaccharide producing mutants the ancestral strain SBW25 was allowed to evolve in a static broth environment for 5 days. The static broth environment is typically 6 ml KB medium contained in a 25 ml microcosm at 28° C. Populations are typically founded from single ancestral ‘smooth’ (SM morph) cells (see Rainey &amp; Travisano, 1998, Nature, 394: 69-72 for illustrations of the principal morph classes of  P. fluorescens  SBW25). Microcosms are incubated without shaking to produce a spatially heterogeneous environment. After 5 days the population shows substantial phenotypic diversity which is easily seen after plating to single colonies on KB agar. During incubation in the static broth culture there is a strong selection for mutants that over-produce glucan-like polysaccharide. These mutants are clearly visible on plating because of their wrinkly spreader colony morphology.  
       Example 2  
       [0217]    Protocol for Polysaccharide Extraction.  
         [0218]    The following protocol may be used for small-scale extraction of the glucan-like polysaccharide of the invention. The protocol as followed up until dehydration will preserve the structure of the polysaccharide network; dehydration or denaturation (by heating or using solvents) may destroy the network. The procedure is designed to first lyse cells, and then to remove proteins through extended proteolysis. Since the procedure involves repeated buffer changes, a lot of the soluble material will be removed from the final sample.  
         [0219]    Colonies:  
         [0220]    1. Grow up WS colonies on hard LB agar plates (2-3 days incubation): 5 μl o/n culture dotted onto a hard (1.5% agar) LB plate, four dots per plate, incubated at 28° C. for 2-3 days (the colonies should be about 20 mm wide before harvest).  
         [0221]    2. Add 2 ml LB broth to plates and leave to stand for 10 min.  
         [0222]    3. Roll off colonies and transfer to a small petri dish, remove broth and transfer to a 5 ml Bejou tube.  
         [0223]    Mats:  
         [0224]    1. Grow standard microcosms (2-3 days incubation): 60 μl o/n culture into 6 ml KB, lid on lightly. Incubate at 28° C. without shaking.  
         [0225]    2. Carefully tip the mat onto a petri dish. Drain off excess liquid and transfer mat to a 5 ml Bejou tube.  
         [0226]    Preparation:  
         [0227]    1. Lyse sample overnight at 37° C. in 5 ml Lysis solution I (with fresh lysozyme)  
         [0228]    2. Remove 4 ml of lysozyme buffer.  
         [0229]    3. Add DNAase and RNAase. Incubate for 30 min at 37° C.  
         [0230]    4. Cool sample on ice for 5 min.  
         [0231]    5. Incubate colony for 24 hr in 5 ml Lysis solution II (with fresh Protease K). Samples should become transparent with trapped air bubbles.  
         [0232]    6. Transfer the glucan-like polysaccharide (GLP) using a P1000 tip cut off at the end (approx. 500 μl) into 5 ml 0.1% SDS for 30 min. at 37° C. Repeat twice more.  
         [0233]    7. Transfer GLP into 5 ml water at RT for five minutes. Repeat four times.  
         [0234]    8. Store at 4° C. until needed.  
         [0235]    EtOH/dehydration:  
         [0236]    1. Wash twice with 50% EtOH for 10 min.  
         [0237]    2. Wash twice with 70% EtOH for 10 min.  
         [0238]    3. Wash twice with 100% EtOH for 10 min.  
         [0239]    4. Transfer GLP into pre-weighed eppendorf tube. Add 1 ml 100% EtOH.  
         [0240]    5. Dry at 80° C. o/n.  
         [0241]    6. Weigh.  
         [0242]    Alternative: Freeze Drying  
         [0243]    1. Freeze dry sample directly.  
                                             Solutions:                                    Lysis Solution I    20 mM Tris · HCl pH 8.0                5 mM MgC12               0.5% Sarkosyl (Sigma)                1 mg/ml lysozyme (fresh)           Lysis Solution II   500 mM EDTA pH 9.0                 1% Sarkosyl                1.5 mg/ml Protease K                      
 
         [0244]    Notes:  
         [0245]    Each mat is approximately 0.4 g wet weight, with about 0.02 g or less dry weight. The lysis step should not be omitted as going straight to the protease incubation results in a very messy mat/colony which is not transparent. The DNase/RNase treatment may not be necessary where the polysaccharide preparation is to be used for analytical purposes, since nucleic acids are unlikely to interfere with polysaccharide assays. At each stage so long as there is an obvious mat/colony or gloopy mass, then dialysis against water or a new buffer is a good way of clearing the polysaccharide of digested material. Before the gloopy mass is moved by pipetting, the sample should be left to cool at 4° C. or on ice (especially if incubations have been at 37° C.). Alternatively, polysaccharide might be by isolated by extraction in 1% SDS and centrifugation: after incubation at 37° C. followed by centrifugation at 12K, the polysaccharide will pellet; boil the pellet in more SDS buffer and re-centrifuge; this time the polysaccharide will be in solution. If the polysaccharide concentration is high enough, cooling of the sample on ice should allow it to form transparent ‘clouds’. Alternatively, the sample could be dried and resuspended in a small amount of water where the polysaccharide should form a gel (the sample should be boiled first and then allowed to set). Once in this form, the SDS can be dialysed out. An important feature of the purification step is that it generates a polysaccharide film.  
       Example 3  
       [0246]    Analysis of a Glucan-Like Polysaccharide  
         [0247]    An example of glucan-like polysaccharide produced from a single type of exopolysaccharide producing strain was subjected to composition and linkage analysis using a range of techniques. The strain used was a wrinkly-spreader strain of  P. fluorescens  isolated using the procedure described in Example 1. It is, however, to be understood that this strain is only one example of the wide range of exopolysaccharide producing strains provided by the invention, each of which may produce polysaccharides which differ slightly in terms of precise structure and composition. The results of analysis of the polysaccharide produced by this strain are therefore not to be construed as limiting to the invention to polysaccharides of this precise structure and composition.  
         [0248]    The polysaccharide was purified using the procedure described in Example 2 and subjected to the following analyses:  
         [0249]    (1) MALDI-TOF Mass Spectrometry  
         [0250]    The polysaccharide material was digested with cellulase and subjected to MALDI-TOF mass spectrometry. A specific peak corresponding to a hexose heptamer was identified by comparison to dextran oligosaccharides.  
         [0251]    (2) Acid Hydrolysis  
         [0252]    The polysaccharide material was subjected to acid hydrolysis which identified glucose as the major sugar residue. Significant amounts of glucose were only released after pre-treatment in 12M sulphuric acid at 25° C. for 30 minutes. This treatment disrupts cellulose fibres, making the individual cellulose chains more susceptible to subsequent hydrolysis when diluted to 0.5M sulphuric acid.  
         [0253]    The results of these analyses strongly suggest that a polymer of glucose linked by β(1-4)-D-glycosidic bonds is the major structural element of the exopolysaccharide produced by a WS strain of  P. fluorescens.  This may be a cellulose or a cellulose-like polymer which, for example, may contain additional branched or modified sugar residues.  
         [0254]    (3) Composition and Linkage Analysis and NMR  
         [0255]    Composition and linkage analysis was carried out on freeze-dried samples of polysaccharide from the WS strain. For compositional analysis, the samples were hydrolyzed using freshly prepared 1M methanolic-HCl for 16 hours at 80° C. The released sugars were derivatized with Tr-Sil and the sample was analyzed by GC-MS using a Sp2330 Supelco column. Myo-inositol was also added to the sample as an internal standard. For linkage analysis, the sample was methylated using the NaOH/Mel method (Ciucanu and Kerek, 1984: Ciucanu, I., and F. Kerek, F. (1984). A simple and rapid method for the permethylation of carbohydrates.  Carbohydr. Res.  131: 209-217.). The methylated sample was hydrolyzed in 2M TFA at 121° C. for two hours and the hydrolyzed carbohydrate was reduced with sodium borodeuteride at room temperature. The product was acetylated using acetic anhydride at 120° C. for three hours. The derivatized sample was then analyzed by GC-MS. Myo-inositol was added to the sample prior to the reduction step as an internal standard.  
         [0256]    NMR analysis used freeze-dried samples. The freeze-dried samples were deuterium-exchanged by repeated evaporation from CD 3 OD and dissolved in 0.5 mL of CD 3 OD. A 1-D proton spectrum was acquired on a Varian Inova 500 MHz spectrometer at 308° K (35° C.). Proton chemical shifts were measured relative to internal TMS standard (d=0.000 ppm).  
         [0257]    Initial chemical analysis of this material showed that the sample contained ˜30% soluble carbohydrates. Of this fraction, composition analysis identified substantial amounts of rhamnose (Rha) and glucose (Glc), as well as trace amounts of fatty acids (Table 1). Linkage data showed two main carbohydrate components, rhamnose and glucose, but N-acetyl glucosamine (GlcNAc), N-acetyl fucosamine (FucNAc) and 3-deoxy-D-amino-2-octulosonic acid (Kdo) were not detected in this experiment. However, these residues are often easily destroyed and are more difficult to observe by linkage analysis than by composition analyses. The sample was treated differently in the composition and linkage analyses (solubilisation and cleavage with HCl/Methanol vs. NaOH/Methanol) and the ratio of rhamnose:glucose identified also varied (0.60 vs. 0.15, respectively). This implies that different fractions of the sample were solubilised by each of these treatments, and that a large fraction of the sample remained insoluble. Finally, the  1 H NMR indicated the presence of alkylated groups in the 1-3 ppm and 4-4.2 ppm regions. The signal from the 1-3 ppm region confirms the fatty acids detected by the composition data. In conclusion, the  1 H NMR data correlated well with the composition data showing the presence of fatty acids and carbohydrates.  
                                         TABLE 1                                   Residue   % present                                        Compositional Analysis: 1                 Rhamnose (Rha)   28.7           Xylose (Xyl)   1.5           3-deoxy-D-amino-2-octulosonic acid (Kdo)   6.5           Glucose (Glc)   48.0           N-acetyl glucosamine (GlcNAc)   7.9           N-acetyl fucosamine (FucNAc)   7.4           Linkage Analysis: 2             Terminal Rhamnose (t-Rha)   2.3           2-Rhamnose (2-Rha)   6.7           3-Rhamnose (3-Rha)   3.9           Terminal Glucose (t-Glc)   4.9           4-Glucose (4-Glc)   82.2           2,4 Glucose (2,4-Glc)   Trace           4,6 Glucose (4,6-Glc)   Trace                      
 
         [0258]    1. The sample also showed some β-OH C10:0, β-OH C12:0, C16:0 and C16:1 fatty acids.  
         [0259]    2. The linkage data only showed two main carbohydrate components, Rha and Glc; GlcNAc, FucNAc and Kdo were not detected in this experiment. These residues are often easily destroyed and are more difficult to observe by linkage analysis.  
         [0260]    Total Carbohydrate Analysis  
         [0261]    The following analysis was carried out in order to look at the variation of polysaccharide material from a number of different variants of  P. fluorescens  (SM, WS, WS-6, WS-18 and JB01).  
         [0262]    Biofilm material from 48 SM, WS, WS-6, WS-18 and JB01 microcosms were extracted to isolate total carbohydrate for analysis. WS-6 and WS-18 are strains which express unmodified glucan-like polysaccharide, SM does not express the glucan-like polysaccharide, JB01 expresses glucan-like polysaccharide. In general, the composition of all five samples looked very similar, and in each, Rha, Kdo, Glc, GalNAc (N-acetyl galactosamine), GlcNAc and FucNAc were identified (Table 2).  
                                                                                       TABLE 2                           Mole (%) and Ratio (to Kdo)            Residues   SM   WS   WS-6   WS-18   JB01                    Rha   11.5   1.69   14.6   4.56   12.2   3.13   17.2   5.06   23.9   5.43       Kdo   6.8   1.00   3.2   1.00   3.9   1.00   3.4   2.00   4.4   1.00       Glc   40.3   5.93   58.4   18.25   51.8   13.28   50.1   14.74   35.1   7.98       GalNAc   2.2   0.32   0.8   0.25   3.0   0.77   1.9   0.56   3.2   0.73       GlcNAc   24.8   3.65   13.3   4.16   18.0   4.62   18.0   5.29   18.1   4.11       FucNAc   14.4   2.12   9.7   3.03   11.1   2.85   9.2   2.71   15.3   3.48                  
 
         [0263]    The overall conclusion from the structural and compositional analysis was that the glucan-like polysaccharide was predominantly a substituted β (1-4) glucan. Furthermore, it appears that the degree and type of substitution can vary as the different bacterial variants gave rise to polysaccharide analyses that, while consisting of predominantly the same residues, gave different ratios of the sugar monomeric constituents.  
       Example 4  
       [0264]    WspR Overexpression  
         [0265]    (1) Over-Expression of wspR-12 Causes GLP Production to be Switched on, but Requires NaCl:  
         [0266]    To over-express the gene wspR-12 was amplified by PCR from the SBW25 genome and a ribosome binding site (GAGGA) added 9 nucleotides from the ATG start of the open reading frame. This was then cloned into plasmid pVSP61 (gift from Steve Lindow, Berkeley) where the wspR gene was expressed from a constitutive Plac promoter. When wspR-12 was over-expressed in ancestral SBW25 no effects on GLP production were noted, however, it was observed that GLP production can be triggered by plating SBW25 containing over-expressed wspR-12 onto LB (No GLP production on KB). The critical ingredient addition was found to be NaCl. The NaCl-dependent effect is only observed in SBW25 containing over-expressed wspR-12.  
         [0267]    (2) Variant Alleles of wspR have Radical Effects on GLP Production:  
         [0268]    Several alleles of wspR which have radically different effects on GLP production were produced by PCR-mutagenesis. These radical effects were seen following over-expression of the variant allele in either the ancestral SM genotype or the evolved WS genotype (see above for details of over-expression). WspR-19, when over-expressed in the ancestral (non-GLP producing) genotype, results in constitutive, signal-independent, production of GLP. This allele is highly significant in terms of GLP production. The ancestral genotype produces little or no GLP during ordinary growth, but by introducing wspR-19 in the cell a GLP-over-producing strain is generated. There is the potential to use this a gene to activate GLP production in other Pseudomonas strains that are capable of producing GLP, but fail to do so in the laboratory. The mutation in wspR-19 is at the very end of the N-terminal domain. WspR-14 has a similar effect, but the effect is not as strong as wspR-19. The mutation in wspR-14 is in the linker region. The remaining wspR alleles all have dominant-negative effects on GLP production, i.e., in a WS genotype they switch OFF GLP production. All these mutations are in the C-terminal domain.  
         [0269]    Summary of wspR alleles:— 
         [0270]    wspR-5: Switches GLP production OFF when over-expressed in a GLP-producing WS genotype.  
         [0271]    wspR-9: Switches GLP production OFF when over-expressed in a GLP-producing WS genotype.  
         [0272]    wspR-12: (wild-type) Over-expression causes GLP production to be switched on, but requires NaCl.  
         [0273]    wspR-13: Switches GLP production OFF when over-expressed in a GLP-producing WS genotype.  
         [0274]    wspR-14: Switches GLP production ON when over-expressed.  
         [0275]    wspR-19: Switches GLP production ON when over-expressed.  
       Example 5  
       [0276]    Assays for Inhibitors of Glucan-Like Exopolysaccharide Production, Bacterial Attachment and Biofilm Development  
         [0277]    There are a number of variations possible on assays that can be used to screen for chemicals which will inhibit bacterial attachment, biofilm development and polysaccharide production. In essence, they all rely on the production of glucan-like polysaccharide by WS or a modified SM strain (glucan-like polysaccharide production is regulated by the same factor which regulates attachment; attachment and biofilm development in SBW25 requires glucan-like polysaccharide production).  
         [0278]    Below are the descriptions of the two basic assays, one for agar plates and the other for liquid cultures:  
         [0279]    Assay 1: Agar Plate Assay for Polysaccharide Production.  
         [0280]    1. Prepare overnight cultures of WS and SM.  
         [0281]    2. Dot 5 pl aliquots of the overnight WS and SM cultures onto agar plates containing 0.001% Congo Red plus the test chemical at various concentrations (including one plate with no test chemical as a control). Incubate overnight at 28° C.  
         [0282]    3. Score glucan-like polysaccharide production by uptake of Congo Red stain by the WS colony. If it is red, glucan-like polysaccharide production has not been affected. If it is white, glucan-like polysaccharide production has been preveneted. The SM colony should be white; if the colony has not developed, then the test chemical at the concentration used is toxic to the growth of the bacteria.  
         [0283]    Assay 2: Liquid Broth Assay for Attachment and Biofilm Production  
         [0284]    1. Prepare overnight cultures of WS and SM.  
         [0285]    2. Add 100 pl of overnight WS and SM cultures to 30 ml glass vials containing 6 ml KB broth (standard microcosms described by Rainey and Travisano, 1998 [Rainey, P. B. and Travisano, M. (1998). Adaptive radiation in a heterogeneous environment. Nature 394: 69-72.] plus the test chemical at various concentrations (including one set with no test chemical as a control). Incubate at 28° C. without shaking for 24-48 hours.  
         [0286]    3. Score attachment and biofilm development:  
         [0287]    a. Visually inspect the SM vials. If the test chemical is not toxic, the SM cultures should be cloudy and the meniscus region should be marked with a faint grime or ring (the control vial with SM should be cloudy). If the broth is clear, then there has been no growth of the culture and the test chemical at the concentration used is toxic to the growth of the bacteria.  
         [0288]    b. Visually inspect the WS vials. The control vial with WS should contain an obvious biofilm (mat) floating at the top of the vial connected to the glass walls. If the test chemical affects cellulose production, then the mat should be reduced in volume and thickness compared to the control WS vial. If the test chemical prevents cellulose production, then there will be no biofilm present at all.  
         [0289]    c. In order to test for attachment, follow this procedure:  
         [0290]    i. Swirl the vial and quickly tip all material out of the vial.  
         [0291]    ii. Add 6 ml KB both back into the vial, replace lid and shake vigourously for 30 sec. before tipping out liquid.  
         [0292]    iii. Repeat (ii) twice more. Allow vials to train upside down.  
         [0293]    iv. Visually inspect the vials in the region of the original meniscus. A faint grime mark should be visible for the SM control vial, and a more obvious ring for the WS control (the meniscus is the site of bacterial attachment to the glass vial). Compare these rings with the test chemical vials to determine the effect of the chemicals on attachment.  
         [0294]    v. The visual inspection can be quantified using Crystal Violet staining:  
         [0295]    a. Add 1 ml 0.1% Crystal Violet solution to each vial and roll so that the dye covers the meniscus region for two minutes. Allow the vials to sit for a few minutes then remove all liquid from the bottom using a pipette.  
         [0296]    b. Add 2 ml water to each vial and roll the vial to was the stained region. Allow the water to drain to the bottom and remove with a pippette.  
         [0297]    c. Repeat (b) twice. Allow the vials to drain upside down.  
         [0298]    d. Add 2 ml 90% EtOH to each vial, replace the lid and vortex or shake vigourously untill all stain is washed from the glass.  
         [0299]    e. Remove EtOH/stain and spin at 13K to pellet any debris. Determine the optical density at 570 nm of, the liquid (OD570), and compare against appropriate standards (Crystal violet in 90% EtOH).  
         [0300]    f. Samples from vials with high OD570 readings indicate that significat bacterial attachment has taken place. Low values indicate poor or no attachment.  
         [0301]    Construction/Isolation of Bacterial Strains:  P. fluorescens  wspA-FΔ:  
         [0302]    This strain was constructed using a standard allelic-replacement technique. The polymerase chain reaction (PCR) was used to amplify two pieces of DNA from the chromosome of  P. fluorescens . The upstream fragment ‘A’ included sequences upstream of the wsp operon, and extended to the beginning of the operon, so that it included the promoter and start codon of wspA. The downstream fragment ‘B’ began with the start codon of wspR and finished downstream of the end of wspR. The two PCR fragments were annealed using strand-overlap extension PCR(SOE-PCR) in such a manner that the start codon of wspA in fragment ‘A’ was joined to the start codon of wspR in fragment ‘B’ (so that the wsp promoter and wspA start codon were now in frame with the wspR coding sequence). The new ‘A-B’ fragment was cloned into a suitable suicide vector, and then transferred into  P. fluorescens  SBW25 strains SM (the wild type strain) and WS (the wrinkly spreader strain). Appropriate co-integrant strains were isolated using the antibiotic resistance of the suicide plasmid. These strains were allowed to grow in the absence of selection to allow a second recombination event to occur and the resultant loss of the suicide vector and original wsp sequences. The presence of the new wspA-FΔ sequence was determined by PCR. These strains have lost wspA-F, but still retain and express wspR. Strains:  P. fluorescens  SM wspA-FΔ  P. fluorescens  WS wspA-FΔ.  
         [0303]    [0303] P. fluorescens  wspΔ:  
         [0304]    This strain was constructed using the SOE-PCR method described above for  P. fluorescens  wspA-FΔ, except in this case, all of the wsp sequences were deleted. Fragment ‘B’ included only sequences after the end of the wspR coding sequence. Strains:  P. fluorescens  SM wspΔ  P. fluorescens  WS wspΔ.  
         [0305]    [0305] P. aeruginosa PA 01 wspD:  
         [0306]    This strain was constructed as for  P. fluorescens  wspΔ except that the wsp sequences used to design the PCR primers were all derived from the  P. aeruginosa  PA01 genome (using the wsp-like operon sequence; available from public databases), not the sequences from  P. fluorescens.    
         [0307]    Complete Sequence Annotation for the  P. fluorescens  wss Operon.  
         [0308]    Set out below is a general description of the genes identified in the wss operon sequence illustrated in FIG. 2 (SEQ ID NO:1). The wss operon encoding the cellulose biosynthetic genes and associated genes, is located approximately between 2,200-18,000 bp of the sequence shown in FIG. 2 (SEQ ID NO:1). The gene co-ordinates for the nine genes (wssA-J) of the operon are listed below. For each gene the positions of the first potential start codon (ATG or GTG [M, methionine or V, valine. N.B. Translation might begin from a GTG codon. However, the first residue would still be a methionine. In order to easily place the peptide sequences with the DNA sequences, GTG-encoded start codons have been left as ‘V’ in the peptide sequences] and the first in-frame stop codon are given. The first potential ATG start codon is also marked (M). A comment about the closest known homologue to each gene is given, along with the probable role of each protein.  
         [0309]    2251-3478 wssA Coding Region (Including Upstream Elements).  
         [0310]    2251-2358 Upstream region; rich in runs of A.  
         [0311]    2309-2358 Predicted promoter region for wssA (5′-CATCAAAATA CTGACACCAT CATTGTGATA TCACAGAATG AGCCCGACAC-3′; A is the predicted transcription start site).  
         [0312]    2425-2430 Possible ribosome binding site (RBS) (5′-AGGTGG-3′).  
         [0313]    2444-2446 GTG start codon: this is the first possible start codon for WssA. This would produce a protein of 344 amino acids. This start codon is preferred to the first in-frame ATG codon on the basis of  E. coli  YhjQ homology. WssA is a MinD homologue and contains an ATP-binding motif.  
         [0314]    2876-2878 ATG start codon: this is the first in-frame ATG start codon for WssA. This would produce a protein of 200 amino acids.  
         [0315]    3476-3478 TGA stop codon: end of WssA.  
         [0316]    3475-5694 wssB Coding Region.  
         [0317]    3475-3477 ATG start codon: this is the first possible start site for WssB. This would produce a protein of 739 amino acids. WssB is a  A. xylinus  BcsA (cellulose synthase A subunit) and  E. coli  YhjO homologue.  
         [0318]    5692-5694 TGA stop codon: end of WssB.  
         [0319]    5884-7953 wssC coding region.  
         [0320]    5884-5886 GTG start codon: this is the first possible start codon for WssC. This would produce a protein of 689 amino acids. WssC is a  A. xylinus  BcsB (cellulose synthase B subunit) and  E. coli  YhjN homologue.  
         [0321]    6148-6150 ATG start codon: this is the first in-frame ATG start codon for WssC. This would produce a protein of 601 amino acids.  
         [0322]    7951-7953 TGA stop codon: end of WssC.  
         [0323]    7884-9146 wssD Coding Region.  
         [0324]    7884-7886 GTG start codon: this is the first possible start codon for WssD. This would produce a protein of 436 amino acids. WssD shares homology with D-family cellulases often found associated with cellulose synthases. WssD is a  A. xylinus  CMCase and  E. coli  YhjM homologue.  
         [0325]    7950-7952 ATG start codon: this is the first in-frame ATG start codon for WssD. This would produce a protein of 398 amino acids.  
         [0326]    9144-9146 TAG stop codon: end of WssD.  
         [0327]    9128-12967 wssE coding region.  
         [0328]    9128-9130 ATG start codon: this is the first possible start codon for WssE. This would produce a protein of 1279 amino acids. WssE is a  A. xylinus  BcsC (cellulose synthase C subunit) and  E. coli  YhjL homologue.  
         [0329]    12965-12967 TGA stop codon: end of WssE.  
         [0330]    12984-13649 wssF Coding Region.  
         [0331]    12984-12986 ATG start codon: this is the first possible start codon for WssF. This would produce a protein of 221 amino acids. WssF is a  A. xylinus  BcsX homologue, required for cellulase expression.  
         [0332]    13647-13649 TGA stop codon: end of WssF.  
         [0333]    13649-14314 wssG coding region.  
         [0334]    13649-13651 ATG start codon: this is the first possible start codon for WssG. This would produce a protein of 221 amino acids. WssG is a  P. aeruginosa  AlgF homologue, required for the acetylation of alginate.  
         [0335]    14312-14314 TGA stop codon: end of WssG.  
         [0336]    14332-15738 wssH Coding Region.  
         [0337]    14332-14334 ATG start codon: this is the first possible start codon for WssH. This would produce a protein of 468 amino acids. WssH is an  A. vineladii  AlgI and  P. aeruginosa  AlgI homologue, required for the acetylation of alginate.  
         [0338]    15736-15738 TAA stop codon: end of WssH.  
         [0339]    15751-16875 wssI coding region.  
         [0340]    15751-15753 ATG start codon: this is the first possible start codon for WssI. This would produce a protein of 374 amino acids. WssI is an  A. vineladii  AlgV/X and  P. aeruginosa  AlgJ/X homologue, required for the acetylation of alginate.  
         [0341]    16873-16875 TAA stop codon: end of WssI.  
         [0342]    16938-17912 wssJ Coding Region.  
         [0343]    16938-16940 GTG start codon: this is the first possible start codon for WssJ. This would produce a protein of 324 amino acids.  
         [0344]    17052-17054 ATG start codon: this is the first in-frame ATG start codon for WssJ. This would produce a protein of 286 amino acids. WssJ is a WssA homologue, and contains an ATP-binding motif.  
         [0345]    17910-17912 TAG stop codon: end of WssJ.  
         [0346]    17913-20306 Downstream region of the wss operon (end of operon, no further significant coding regions).  
         [0347]    Complete Sequence Annotation for the  Pseudomonas fluorescens  wsp Operon  
         [0348]    Set out below is a general description of the genes identified in the wsp operon sequence illustrated in SEQ ID NO: 27. The sequence shown as SEQ ID NO: 27 is contiguous piece of DNA of 13,288 bp from  Pseudomonas fluorescens  SBW25. The wsp operon encodes a chemotaxis-like operon of seven genes, wspA-F and wspr. A schematic arrangement of the operon is shown as FIG. 28.  
         [0349]    The gene co-ordinates for the seven genes of the operon are listed below. For each gene the predicted polypeptide sequence is given, from the first potential start codon [ATG encoding M, methionine] to the first in-frame stop codon. A comment about the closest homologue to each gene is given, along with the probable role of each protein.  
         [0350]    4535-6178 wspA Coding Region.  
         [0351]    4535-4537 ATG start codon: this is the first possible start codon for WspA. This would produce a protein of 547 amino acids. WspA is a MCP homologue. The deduced amino acid sequence of WspA exhibited significant similarity to methyl-accepting chemotaxis proteins (MCPs), sensory transducers involved in bacterial chemotaxis and motility. The highest similarity was shared with the probable chemotaxis transducer PA3708 from Pseudomonas aeruginosa PA01 (accession C83184) (E-value=1e-167; Identities 60%; Positives 70%) and with a chemotaxis transducer from the plasmid pMLb of  Mesorhizobium loti  (accession NP — 109390) (E-value=1e-120; Identities 45%; Positives 59%).  
         [0352]    6176-6178 TGA stop codon: end of WspA.  
         [0353]    6178-6690 wspB Coding Region.  
         [0354]    6178-6180 ATG start codon: this is the first possible start site for WspB. This would produce a protein of 170 amino acids. WspB is a CheWI homologue. The deduced amino acid sequences of WspB is similar to the chemotactic protein CheW, involved in the transmission of sensory signals from bacterial chemoreceptors (MCPs) to the regulatory components that control chemotaxis and motility. WspB shared highest similarity with the hypothetical protein PA3707 from  Pseudomonas aeruginosa  PA01 (accession B83184) (E-value=1e-43; Identities 60%; Positives 71%). Similarity was also high with a hypothetical protein from the plasmid pMLb of  Mesorhizobium loti  (accession NP — 109389) (E-value=6e-28; Identities 42%; Positives 61%), with the chemotaxis protein CheW from  Rhizobium meliloti  (accession Q52881) (E-value=5e-07; Identities 28%; Positives 50%) and other CheW-like bacterial proteins.  
         [0355]    6688-6690 TGA stop codon: end of WspB  
         [0356]    6687-7946 wspC Coding Region.  
         [0357]    6687-6689 ATG start codon: this is the first possible start codon for WspC. This would produce a protein of 419 amino acids. WspC is a CheR homologue. The deduced amino acid sequence of WspC exhibited significant similarity to the chemotaxis protein CheR, belonging to the superfamily of protein-glutamate O-methyltransferases involved in the methylation of MCPs; the methylation state of the MCPs in the cell is crucial for sensory responses and adaptations. The highest similarity was shared with the probable protein methyltransferase PA3706 from  Pseudomonas aeruginosa  PA01 (accession A83184) (E-value=1e-116; Identities 57%; Positives 66%) and with a methyltransferase from the plasmid pMLb of  Mesorhizobium loti  (accession NP — 109388) (E-value=1e-71; Identities 38%; Positives 52%).  
         [0358]    7944-7946 TGA stop codon: end of WspC.  
         [0359]    7943-8641 wspD Coding Region.  
         [0360]    7943-7945 ATG start codon: this is the first possible start codon for WssD. This would produce a protein of 232 amino acids. WspD is a ChewII homologue. The deduced amino acid sequences of WspD is similar to the chemotactic protein CheW, involved in the transmission of sensory signals from bacterial chemoreceptors (MCPs) to the regulatory components that control chemotaxis and motility. WspD only shared high similarity with the hypothetical protein PA3705 from  Pseudomonas aeruginosa  PA01 (accession H83183) (E-value=7e-58; Identities 55%; Positives 67%), with a hypothetical protein from the plasmid pMLb of  Mesorhizobium loti  (accession NP — 109387) (E-value=1e-41; Identities 42%; Positives 56%) and with the chemotaxis protein CheW from  Rhodobacter sphaeroides  (accession Q60251) (E-value=2e-04; Identities 24%; Positives 42%).  
         [0361]    8639-8641 TAG stop codon: end of WspD.  
         [0362]    8638-10905 wspE Coding Region.  
         [0363]    8638-8640 ATG start codon: this is the first possible start codon for WspE. This would produce a protein of 750 amino acids. WspE is a CheA homologue. The deduced amino acid sequence of WspE exhibited similarity to hybrid proteins consisting of the chemotactic histidine kinase CheA and the chemotactic response regulator CheY, both involved in the transmission of sensory signals from the chemoreceptors (MCPs) to the flagellar motors. Following autophosphorylation, the histidine kinase CheA transfers its phosphate group to CheY (or the methylesterase CheB), which in turn interacts directly with the flagellar motor controlling chemotactic behaviour and motility. The highest similarity was shared with the probable chemotaxis sensor/effector fusion protein PA3704 from  Pseudomonas aeruginosa  PA01 (accession G83183) (E-value=0.0; Identities 66%; Positives 74%) and with a chemotaxis histidine kinase from the plasmid pMLb of Mesorhizobium loti (accession NP 109386) (E-value=0.0; Identities 49%; Positives 62%).  
         [0364]    10903-10905 TGA stop codon: end of WspE.  
         [0365]    10902-11912 wspF Coding Region.  
         [0366]    10902-10904 ATG start codon: this is the first possible start codon for WspF. This would produce a protein of 336 amino acids. WspF is a CheB homologue. The deduced amino acid sequence of WspF exhibited similarity to the chemotaxis protein CheB, belonging to the family of protein-glutamate methylesterases, involved in the demethylation of MCPs following phosphorylation of their response regulator domain by the histidine kinase CheA. Highest similarity was shared with the probable methylesterase PA3703 from  Pseudomonas aeruginosa  PA01 (accession F83183) (E-value=1e-138; Identities 73%; Positives 84%) and with a methylesterase from the plasmid pMLb of  Mesorhizobium loti  (accession NP — 109385) (E-value=8e-93; Identities 53%; Positives 67%).  
         [0367]    11910-11912 TGA stop codon: end of WspF.  
         [0368]    11962-12963 wspR Coding Region.  
         [0369]    11962-11964 ATG start codon: this is the first possible start codon for WspR. This would produce a protein of 333 amino acids. WspR is a response regulator and exhibited highest similarity with the probable two-component response regulator PA3702 from  Pseudomonas aeruginosa  PA01 (accession E83183) (E-value=1e-131; Identities 74%;  
         [0370]    Positives 85%) and with a regulatory component from the plasmid pMLb of Mesorhizobium loti (accession NP — 109384) (E-value=2e-98; Identities 57%; Positives 71%). 12961-12963 TAG stop codon: end of WspR.  
       Sequence Listing  
       [0371]    SEQ ID NO: 1  Pseudomonas fluorescens  wss operon, complete nucleotide sequence  
         [0372]    SEQ ID NO: 2 WssA polypeptide sequence  
         [0373]    SEQ ID NO: 3 WssB polypeptide sequence  
         [0374]    SEQ ID NO: 4 WssC polypeptide sequence  
         [0375]    SEQ ID NO: 5 WssD polypeptide sequence  
         [0376]    SEQ ID NO: 6 WssE polypeptide sequence  
         [0377]    SEQ ID NO: 7 WssF polypeptide sequence  
         [0378]    SEQ ID NO: 8 WssG polypeptide sequence  
         [0379]    SEQ ID NO: 9 WssH polypeptide sequence  
         [0380]    SEQ ID NO: 10 WssI polypeptide sequence  
         [0381]    SEQ ID NO: 11 WssJ polypeptide sequence  
         [0382]    SEQ ID NO: 12 WspR-12 polypeptide sequence  
         [0383]    SEQ ID NO: 13 WspR-12 nucleotide sequence  
         [0384]    SEQ, ID NO: 14 WspR-5 polypeptide sequence  
         [0385]    SEQ ID NO: 15 WspR-5 nucleotide sequence  
         [0386]    SEQ ID NO: 16 WspR-9 polypeptide sequence  
         [0387]    SEQ ID NO: 17 WspR-9 nucleotide sequence  
         [0388]    SEQ ID NO: 18 WspR-13 polypeptide sequence  
         [0389]    SEQ ID NO: 19 WspR-13 nucleotide sequence  
         [0390]    SEQ ID NO: 20 WspR-14 polypeptide sequence  
         [0391]    SEQ ID NO: 21 WspR-14 nucleotide sequence  
         [0392]    SEQ ID NO: 22 WspR-19 polypeptide sequence  
         [0393]    SEQ ID NO: 23 WspR-19 nucleotide sequence  
         [0394]    SEQ ID NO: 24 pgi nucleotide sequence  
         [0395]    SEQ ID NO: 25 mreB nucleotide sequence  
         [0396]    SEQ ID NO: 26  E. coli  chromosomal sequence, including the yhj operon  
         [0397]    SEQ ID NO: 27  Pseudomonas fluorescens  Wsp operon, complete nucleotide sequence  
         [0398]    SEQ ID NO: 28 WspA polypeptide sequence  
         [0399]    SEQ ID NO: 29 WspB polypeptide sequence  
         [0400]    SEQ ID NO: 30 WspC polypeptide sequence  
         [0401]    SEQ ID NO: 31 WspD polypeptide sequence  
         [0402]    SEQ ID NO: 32 WspE polypeptide sequence  
         [0403]    SEQ ID NO: 33 WspF polypeptide sequence  
         [0404]    SEQ ID NO: 34 WspR polypeptide sequence (wild-type)  
     
       
       
         1 
         
           
             34  
           
           
             1  
             20306  
             DNA  
             Pseudomonas fluorescens  
           
            1 

ggtaccaaca tcgacgacgg caccaaccgc ggcgatggct ccgagcgtga aatctggaac     60 

cagttcaagt acgtggtcca gagcggtccg gccaaagacc tgagcctgcg tgctcgtgcc    120 

tcgtggctgc gcgtctccaa taacgctgac cagtacaacg taggcggtaa cgaaatccgt    180 

ctgttcgccg actacccgat caacgttttc taattgatct ggcgtcgcgc ctgatgccag    240 

gcgctaaaaa accccgactg gttcggggtt tttttatgcc cgccagtgcg agacacgttt    300 

gttacaccac ggcactgatg atgtacccgg ccccaccttt atcccctccc ggcgcatgcc    360 

tacggcttct tctatatccc ccaactcgcc cttgccgaga tggaaaagaa caacgccggc    420 

cgcatcgtcc acatcaccac cagcctggct gaccatgcaa tcgatggcgt accgtcggta    480 

cttgccaacc tcaccaaagg cggcctgaac tcagcgccca aatcattggc gattgaatac    540 

gccaagcgcg gtatccgagt gaatgcggta agcccgggga tcatcaagac gccgatgcat    600 

ggcgaggaaa gccatgctgc gctggggcaa atgcacccgg ttgggcacat gggggaggtg    660 

agtgaaatcg cccaggcgat tatttatctg gagaatgcgg ggttcgtgac gggggagatt    720 

cttcatgtag acggcgggca gagtgccggg cattgatcta cacgtcccgg cttgattggg    780 

ccgggcgtta tcttgtcttg gtgaggttgt gcgtaggctg ggaggggggc acctgccatt    840 

tggtacttta agaaattgag ctcttccggg aaataggcag aaggtgacaa ggtcggagaa    900 

gctatcaacc aggtgagggc gtcggttgag caaaaatgaa gcgtggacac tttttgggca    960 

ttttggccaa aacgacgccc aagatttttc gtgttttcca aacccaagaa accacaaagc   1020 

cccgcattgc ggggcttcga gatatggtgc cggcaccagg agtcgaaccc gggacctact   1080 

gattacaagt cagttgctct accaactgag ctataccggc gtgttagggc gacgattata   1140 

gcgattggaa aggttctgta aacccctgaa ttctgactat ttttgcaaaa cccaagcttt   1200 

tctcacactc gccccttttt tcgaccagca agggtagatt cttctcttac tgccgaaggg   1260 

tctaactcgg caatcttgtc caaactaaag gaagccccat atgaaacgga ctctctccct   1320 

gtccctcgtc ctcctcaccg ccgctctcgg cgcgtgctcc acccatcagt cggccaacga   1380 

ccccgcgctg gttggcactt ggaaaggctt gcgcaccgag accggtaaat gccagttcct   1440 

gtcgtggacc aataccctca agcctgacgg tcgtttcgtt attaccttct accgcgatgc   1500 

gcagcagacg caggtgatcc acacggacac ggttcgtggg cggcggccaa tggttagaat   1560 

gagctgcgca cagatcgcgt gcgttcgccg gatgtgttca cctataagct gctggacgca   1620 

gacactgttc actatgtgag tgtggcatcg gaccccacca gcgattgcca ggacgactac   1680 

cagttcaccg agcgccgtgt ccgctgacaa ggcaggtttg cttcaaaacc gggcactcgg   1740 

gtgcccggcc gtcacaagtt tcaactccct tgcgcgcacg tgcccgcacc tttctcgtag   1800 

tccaatacct gtttttgccg ggcgtgagca tcagcagcaa aagcctacag cctgactgcg   1860 

atcaatttgg tctagttaat ggggcgtatc ttccaggctc agcaggttta gcgctctgga   1920 

ggtgtctgcc ctgtgtcatt cggcgtgggt tttgcttcgt cgtcttggct gcctgcgcag   1980 

cgatatacgg gccgtggctt tgcgggcggt tgcggatcga ccgctcggca gtttcgtcct   2040 

tgaatacagg cgaggtttcc gtattctgtc gcgcgatata aaccgtccgt ttaatcgcgg   2100 

ttgtgtgtag gcaaattttt gagacagtgt agggcaatcg aatatttgtt tatgggtatg   2160 

tcagattagt gccgtttctg accactcagc caacaaataa tacgataaat cagtgcaagt   2220 

tcatgttctt acagcgaatg cattctctct aaataatctg tcataaaaat ggcgccacgc   2280 

cgttcgtcgg ataagattga gcgaaaagca tcaaaatact gacaccatca ttgtgatatc   2340 

acagaatgag cccgacacca cataactaaa aaaatacaag gtcagcctat tgctgacctt   2400 

tttatcgcct ttaaagcgca aaccaggtgg ggaggggcag tgagtgagtc gagcagatga   2460 

catttcaaaa ctattcaaca agctgggtgc caacccgagt ggctaccgcg agatcgactt   2520 

cgtccacgag ttcatcgagg acgatgtaga ggtcctggaa accccggcgg tcgttcgagc   2580 

cctgcctgtg attgaagcgc cgtcagcgcc tttgctgcgt ctgttggaag agctgagcca   2640 

aggcgaagcc gaccacctgc agccgcccga agtggtggaa gggcgggacg gtgaggtgta   2700 

ctcggagcat tccagcccca acgtcgtggt ggttgtctcg gtaaaaggcg gcgttggccg   2760 

cagcaccctg actgccgcga ttgccagtgg tttgcagcgt caggggcgcc cggcactggc   2820 

cctggacctg gacccgcaaa acgccctgcg ccaccacttg tgcctcggtc tcgacatgcc   2880 

cggcgtgggc gcgaccagct tgctcaatga aagctgggaa gcgctgcccg agcgcggttt   2940 

tgccgggtgc cgcctggtgg cattcggtgc taccgaccac gagcagcaac agagcctgaa   3000 

tcgctggctg ggccaggatg acgaatggct gagcaaacgc ttggccggcc tcaaattgaa   3060 

cggccaggac accgtgatca tcgacgtccc ggccggcaat accgtgtact tcagccaagc   3120 

catgtcggtc gccgacgcag tgctggtagt ggtgcagccg gacgtggcgt ccttcagcac   3180 

gctcgatcag atggacagcg tgctcaagcc ctctctcaat cgcaaaaaaa cgccacgacg   3240 

cttctatgtg atcaaccaac tggacggtgc ccaccgcttc agcctggaca tggccgaggt   3300 

gttcaagacc cgcctgggtg ctgccctgtt ggggacggtc caccgcgacc ccgcgttcag   3360 

cgaagcccag gcctacgggc gtgatcccct tgaccccacc gtcaacagta tcggcagcca   3420 

ggacatccat gccctgtgcc gcgcattgct cgaacgaatc gactcggacc tcccatgacc   3480 

gacactacgt cctccacgcc cttcgtcgaa gggcgcgctg aacagcgcct aaatggcgcc   3540 

atcgcgcgct tcaatcggtg gccttcggcg ccgcgtacgg tactggttgt tgccagttgc   3600 

gtgctcggcg ccatgctgct gctgggcatt atcagcgcgc cgctcgacct ctacagccaa   3660 

tgcctgtttg ccgccgtgtg cttcctggcg gtgctggtac tgcgcaagat cccggggcgc   3720 

ctggcgatcc tcgccttggt ggtgttgtcg ctggtggcgt cgttgcgcta catgttctgg   3780 

cgcctcacct ccaccctcgg ctttgaaacc tgggtcgaca tgttcttcgg ctacggcctg   3840 

gtcgcggccg agttctacgc cctgatcgtg ctgatcttcg gctacgtgca aaccgcctgg   3900 

ccgctgcgcc gcacgccggt gtggctcaag actgagccgg aagagtggcc gacggtcgac   3960 

gtgttcatcc cgacgtataa cgaggcgctg agcatcgtaa agctgaccat tttcgccgcc   4020 

caggcgatgg actggcccaa ggacaagctg cgcgtccacg tgctcgacga cggtcgccgc   4080 

gacgacttcc gcgaattctg ccgcaaggtc ggcgtgaact acatccggcg cgacaataac   4140 

ttccacgcca aggccggtaa cctcaacgaa gcgttgaagg tcaccgacgg cgaatacatc   4200 

gccctgttcg acgccgacca cgtgccgacg cgttccttcc tgcaagtgag cctgggctgg   4260 

ttcctgaaag atccgaagct ggcgatgctg caaacgccgc acttcttctt ctcgccggac   4320 

ccgtttgaaa agaacctcga cacgttccgc gccgtgccca acgaaggtga gctgttctac   4380 

ggcctggtgc aggacggcaa cgacctgtgg aacgccacct tcttctgtgg ctcctgtgcc   4440 

gtgattcgcc gtgagccgct gctggaaatc ggcggcgtag ccgttgagac cgtgaccgaa   4500 

gacgcccaca ccgcgctcaa gctcaaccgc ctgggctaca acaccgctta cctggccatc   4560 

ccacaagccg cgggcctggc cactgaaagc ctgtcgcgcc acatcaacca gcgcattcgc   4620 

tgggcacggg gcatggcgca gattttccgc accgacaacc cgctgctggg caaaggcctg   4680 

aagtggggcc agcgcatctg ttatgccaac gccatgcagc acttcttcta tggtttgccg   4740 

cgcctggtgt ttttgaccgc gccgttggcc tacctgattt tcggcgccga aatcttccac   4800 

gcatcggcgc tgatgatcgt cgcctatgtg ttgccgcact tggtgcactc cagcctgacc   4860 

aactcgcgga tccaggggcg cttccgtcac tcgttctgga acgaggtgta cgagaccgta   4920 

ctggcctggt acatcctgcc gccggtgctg gtcgcgctgg tcaatcccaa ggcgggtggc   4980 

ttcaacgtca ccgacaaggg cggcatcatc gacaagcagt tcttcgactg gaaactcgcc   5040 

cggccctacc tggtgttgtt ggcggtcaac ctgatcggcc tcggcttcgg tatccaccag   5100 

ttgatctggg gcgatgcgtc cactgccgtg acggtggcga tcaacctgac ctggaccctc   5160 

tataacctga tcatcaccag tgccgccgtg gccgtggcct cggaagcacg ccaagtgcgt   5220 

tccgagccgc gtgtcagcgc caagctgcca gtgagcatca tttgcgccga cggccgcgtg   5280 

cttgatggca ccacccagga cttctcgcaa aacggcttcg gcttgatgct ttccgatggt   5340 

cactcgatca cccagggtga acgcgtgcaa ttggtactgt cgcgcaacgg ccaggacagc   5400 

ctgaaagacg cccgcgtggt gttcagcaag ggcgcgcaga tcggcgccca attcgaagcc   5460 

ctcagcctgc gccagcaaag cgaactggtt cgcctgactt tctcccgcgc cgatacctgg   5520 

gccgcgagtt ggggcgccgg tcagccggat acgccgctgg cggccctgcg cgaagtcggc   5580 

tccatcggca tcggcggctt gttcaccctc ggccgcgcca ccctacacga actgcgtctg   5640 

gccttgagcc gcacccccac aaaaccgcta gatacgctga tggacaagcc atgacctcga   5700 

acatattcgc tcgccctcat ccgcgccgtg cactggcgct gatgatcgcc tcgctgatgg   5760 

gcttcaacac cctcgcgcaa gctgccgagc aagcggttgc caccgtgccg gtacaaagta   5820 

ccgacaccgg ctacagcctg accctcaagc aattggggcg tcgcgacacc atgaacctgc   5880 

aaggtgtgga gtcgtccaca gcgtaacttt gatatccgtg ccgatgaagt ggtgaagggc   5940 

gcgcaactgt tgctcaagta cagctactcg ccggcgctgc tggccgatct gtcgcagatc   6000 

aacgtgctgg tcaacggcga agtcgccgcc agcctgccgc tgcctaaaga aggcgcgggc   6060 

acgccgcaag agcagttggt gcagatccca gcgcacttga tcaccgagtt caaccgcctg   6120 

agcctgcagt tcatcggcca ctacaccatg tcttgcgaag acccactgca cagcagcctg   6180 

tgggccaaga tcagcaacag cagtgagctg aaagtgcagg tcgagccgat cgtgctcaaa   6240 

gacgacttgg ccgtgctgcc gctgccgttc ttcgacaagc gtgacgcacg ccaggtgagc   6300 

ctgccgttcg tgttcgccac cgctcccgac agcgccgcgc tggaagccgc cggcgctctg   6360 

tcgtcgtgga tcggcggcct cgccagctac cgcggcgcga cgttcccgac caccctcggt   6420 

gagttgccgg ccaaaggcaa cgccatcgtg ctggtgcaaa ctgccgacgc gatggacata   6480 

cacggtgttg ccgttgccaa gccggccggc ccgaccctca ccctcatcgc caacccgaac   6540 

gacgccaacg gcaagctgct gatcgtcacc ggccgtgacg gtgcagagct caagcgcgcc   6600 

gcaacgcggt ggtgctggca acccggtatt ggccggtaca gcgtggtgat caccaagctg   6660 

gataccctcg caccgcgtcg tccatacgat gccccgaact ggctgccaag caaccgcccg   6720 

gtccgcctgg gcgagttgat cgaacagcaa aaactcagcg tgtcgggcta caacccaggc   6780 

gcgatcagcg tcgatatgcg cctgccgcca gacctgttca actggcgtga agagggtgtg   6840 

ccgctcaagc tcaaataccg ctacacgccg cagcaggtgt cgaccaactc gtcgctgctg   6900 

atcggcctga atgatcagtt catgaagtcc gtggcattgc cgtcggtgag caacctgggc   6960 

ggcggccaaa ccctgctcga ccagttgaag aaagacgaaa gcctgccgcg tgaagtgacg   7020 

accctgttgc cgatcagctc agcatcgccc aagtccaagc tgcaagtgcg cttcatgtac   7080 

gactacatca aagaaggcga atgccgcgac atcatcgtcg acaacatgcg cggttcggtc   7140 

gacccggact cgaccctcga cgtcacgggc taccagcact acatcgccat gccgaacctg   7200 

ggcgtgttca acgactcggg tttcccgttc acccgtttgg ctgacctgtc ggagtcggcc   7260 

gtggtcatgc ccgacaatta cggtaccgac gagctgaccg cctacctgac ggtgcttggc   7320 

cgttttggcg aagccaccgg ctacccggcc actgcggtaa aagtagtgca ggccaaagac   7380 

gtgcaaagcg ttgccgacaa agacctgttg gtgctcgcta ccgctgccaa ccaaccgctg   7440 

ctcaagcagt ggcagcaata cctgccggcg accagcgatg gcgagcaaca ccagttcctg   7500 

ctgtctgacc tgccacgtta tgtgcgcagc tggatcagcc cggacccggc ggccaaccaa   7560 

cacccggcca acaccggcat taccttcaag ggcctgagca acagcacgtg gctggcgggt   7620 

ttccaatcgc cgctcaagag cggtcgcagc gtggtgctga tcgccagcaa ccagcctcag   7680 

ggcctgctgg aagcgaccaa cgcgctgatc ggcggtgacg actataaaga ctcgatccaa   7740 

ggcagcctgg cggtggtgca aggcacgcaa atcagctcgc tggtaggcga tgagcagtac   7800 

tacgtcggca agctcaacta cttcaaattc atgcagtggc aactgtcgca gaacctgggt   7860 

tggatgctgt tgatcacctt cctcggcctg gccgtggtca ccagcctgat ctacttgtcg   7920 

ctgcgtgccc gtgcaaaacg gcggttggca tgagcccgct taagtgcatg gccctggcgg   7980 

cgctgggcgc tgtgatgttc gtcggtagcg cgcaagcgca aacctgcgac tggccgttgt   8040 

ggcagaacta cgccaagcgc ttcgtgcagg atgatgggcg cgtgctgaac tcgtccatga   8100 

aacccaccga gagcagctcg gaaggccaat cctatgcgat gttctttgcc ctggtcggca   8160 

acgaccgtgc tagcttcgac aagctgtgga cctggaccaa ggccaacatg tcgggcgccg   8220 

acatcggcca gaacctgccc ggctggttgt ggggcaaaaa agccgataac acgtggggtg   8280 

tgatcgaccc gaactccgcc agcgacgccg acttgtggat ggcctacgcc ttgctcgaag   8340 

cggcacgcgt gtggaatgca ccgcagtacc gtgccgacgc gcaattgttg ctggcaaacg   8400 

ttgaacgcaa cctgatcgtg cgtgtgcctg gcctcggcaa gatgctgttg ccaggcccgg   8460 

tgggctacgt gcacgccggt ggcctgtggc gcttcaaccc gagctatcag gtgctggcgc   8520 

aactgcgtcg cttccacaaa gaacgcccga atgccggctg gaatgaggtg gccgacagca   8580 

acgccaagat gcttgccgat acggccagca accctcacgg cctcgcggcc aactgggtcg   8640 

gttaccgcgc caccagcgcc aacaccgggc tgttcgtggt cgacccgttt tccgatgact   8700 

tgggcagcta tgacgcgatc cgcacttaca tgtgggccgg catgaccgcc aagggcgatc   8760 

cgctggcggc gcccatgttg aaatccctgg gcggcatgac gcgggccacc gcagcatcgg   8820 

ccaccggcta cccaccggaa aaaatccatg tactgaccgg cgaagtcgag aaaaacaacg   8880 

gctatacgcc gatgggcttc tcggcctcca ccgttgcctt cttccaggct cgcggcgaaa   8940 

ccgcactggc gcagttgcag aaggccaagg tcgacgacgc gctcgccaaa gccctggccc   9000 

cttcggcgcc ggacaccgcg cagccgatct actacgacta catgctcagc ctgttcagcc   9060 

agggctttgc cgatcagaag taccgttttg aacaagacgg tacggtcaaa ctttcctggg   9120 

aggccgcatg cgccgtcaca cgctagccat tgcgattctc gccgccctgg cctccaccgc   9180 

cagcgtcgct gaaaccagcg acccgcagtc cctgctgatt gagcagggct actactggca   9240 

gtcgaaaaag aaccccgaac gcgctctgga aacctggcag aaattgctgc gcctgagccc   9300 

ggaccaaccg gatgcgctgt acgggatcgg cctgatccag gtgcaacaga accacccggc   9360 

cgaagcgcaa aagtacctgg cccgtttgca agcgttgagc ccggtgccgc gccaggcctt   9420 

gcaactggaa caggacatca ccgttgcggt tccggacaac gccaagttgc tggaacaggc   9480 

gcgtgaactg ggcgagccgg aggccgagcg tgaacaggct gtggcgttgt atcgccagat   9540 

tttccagggc cgtcagcctc aaggcctgat cgcccgcgag tactacaaca ccctgggctt   9600 

taccgccaaa ggcagcagcg aggccatcgc cggcctgcag cgcctgaccc gtgaacgccc   9660 

gaacgacccg atcgttgcgc tgttcctggc caagcacctg gcgcgtaacc cggccacccg   9720 

gcctgacggt atacgggccc tggctaaact ggcatcgaac aacgacgtgg gcggcaacgc   9780 

tgacgaaacc tggcgcttcg cgctggtctg gctcggcccg ccgaaaccgg accaggtttc   9840 

gctgttccag cagttcctca ccgtgcaccc ggatgacagc gagatccgcg cgctgatgaa   9900 

caaaggcatc gcccaaggta aaggcggtgg cacctggcag cgtgatccgc agatgaccaa   9960 

ggctttcaag gcgttggatg acggcgacct gaaaaccgcc gagcctctgc tggcggcgcg  10020 

ccttgcgcaa aagtccaatg acgttgatgc cctcggcggc atgggcgtgt tgcgtcaaca  10080 

gcaagagcgc tacagcgagg cggaaaacta cctggtccaa gccacgcgct tgccgggtgg  10140 

cgctgcttgg cagtcggcct tgaatgatgt gcgctactgg aatctgatca gccaaagtcg  10200 

tgacgcccag cgtgccggtc gtagcgccca ggcccgcgac ctggtggccc aggccgaacg  10260 

cctgaaccct ggccagcctg gcgcggctat tgcgctggcg ggcttccagg cccaggacaa  10320 

ccagttcgac gacgccgaag cgggctaccg caaagtgctg gctcgccacc ctggcgaccc  10380 

ggatgccttg agcggcttga tcaacgtgct gtcccagtcg ggccagccgg atgaagcctt  10440 

gaagctgatc gactcggtat cgccggccca gcgcgccaag ttcgcaccga gtgtgaaaat  10500 

caacgcgttg cgcgcgaccc aggtgggcaa gctggccgag cagcgggggg atctgaaggc  10560 

cgcgcaagcc gcctatcgcc aggccctcga tgccgacccg gaaaacccct ggacccgctt  10620 

cgccttggcg cgcatgtacc tgcgcgacgg gcagatccgc aacgcgcgag ccttgatcga  10680 

cggcttgctc aagtcccagc ccaaccaacc cgatgcgctg tacaccagca ccttgttgtc  10740 

ggcgcaattg agcgagtgga agcaagccga ggccaccttg gggcgtatcc ccacggcaca  10800 

gcgcacggcg gacatgaacg aactggcaac tgacatcgcg ctgcaccaac agaccgacat  10860 

cgctatcgaa accgctcgcc gtggccagcg cccggaagcc ttggcgttgc tcgggcgttc  10920 

cgaaccgctg acccgcaata aacctgagcg tgtggccgtg ttggccgccg cctatgtgga  10980 

agtgggggct gcgcagtacg gtctggacat gatgcagaag gtggtggaga acaaccccaa  11040 

cccgacggtc gaccagaagc tgctgtacgc caacgtgctg ctcaaggcca acaaatacag  11100 

tgaggcgggc gagatcctgc gcgaagtgca aggccagcct ttgaccgaga ccggccgcca  11160 

gcgctacgac gatttgatct acctgtaccg ggtcaagcag gccgacgccc tgcgtgagaa  11220 

gaacgacctg gtggcggctt acgacatgct gtccccggcc ctggcccagc gcccgaacga  11280 

cgcgttgggc gtcggcgccc tggcgcggat gtacgccgcc agcggtaatg gcaagaaggc  11340 

catggagctg tacgcgccgt tgatccagca gaaccccaat aatgcgcgac tgcaactggg  11400 

ccttgcggac atcgccctga aaggtaatga ccgtggcttg gcgcaaagcg ccagtgacaa  11460 

ggcgttggcc ctggagccgg gtaacccgga gatcctcacc tcggccgcgc gcatctatca  11520 

gggcctgggc aagaacagcg aagccgcgga actgctgcgc aaggccctgg cgattgaaaa  11580 

cgccatgaag gccaagaccc aggtagccca ggccagcgct ccaggtacgt cgtataaccc  11640 

gttcgtgggc ttgccgggcc agcgtcgcca ggtcaccgac ctgaccgttg ccggcgcggt  11700 

accgccaccg atcgatgcac cgaccaagtc cgtgacgtcc aacgcgtttg cgagtgccac  11760 

gtccaacgac ttgagcgacc cgtttgtgcc gccatcgagc attgcgtcga tcgacagccc  11820 

cgagctgagc ccggcgcgtc gtgcgctgga cactatcctg cgtgaccgta ccggttatgt  11880 

ggtgcagggc ttgagcgtac gcagcaacaa cggtgagaaa ggcttgagca aaatcaccga  11940 

tgtggaagcg ccgtttgaag cgcggatgcc ggtgggggat aacactgtgg cgctgcgggt  12000 

gacgccggtg catttgagtg ctggtagcgt caaggctgag tcgttgtcgc gctttggtaa  12060 

aggcggcaca gagccggcgg gctctcagag cgatagcggt gtcggtttgg ccgtggcgtt  12120 

tgaaaacccg gatcaaggcc tcaaggctga cgtcggcgtg agccctctgg gcttcctcta  12180 

caacaccctc gtgggcggcg tgagcgtgtc gcgtccgttc gaggccaatt cgaacttccg  12240 

ctacggcgcc aacatctcgc ggcgcccggt caccgacagc gtcacttcct ttgccggctc  12300 

cgaagacggc gcgggcaaca agtggggggg cgtcaccgcc aacggcggcc gtggcgagct  12360 

gagctatgac aaccagaaac tgggcgtcta cggctacgcg tcgttgcatg agttgctggg  12420 

caacaatgtt gaagacaaca cccgcctgga gttgggcagc ggtatctact ggtacctgcg  12480 

taacaacccg cgcgacacgt tgacgctggg tatcagtggt tcggcgatga ccttcaagga  12540 

aaaccaggac ttctacacct acggcaacgg cggttacttc agcccgcagc gctttttctc  12600 

cctcggcgta ccgattcgtt gggcgcaaag ctttgatcgc ttcagttacc aggtaaaaag  12660 

ttcggtgggc ctgcagcata tcgcgcaaga tggcgcggat tatttccctg gcgacagcac  12720 

gcttcaagcc accaaaaata atccaaagta cgacagcacc agcaagactg gcgtcggcta  12780 

cagcttcaac gccgccgccg aataccgctt gagctcgcgc ttctacctgg gcggcgaaat  12840 

cggtctcgac aacgcccagg actaccgcca gtacgccggt aatgcttatc tgcgctactt  12900 

gttcgaagac ctgagcgggc ctatgccctt gccggtcagt ccgtaccgtt ccccttattc  12960 

caactgatta atcggagtca ttcatgcctg tttctgcgat tgccggccta accatgctgg  13020 

tattgggcga aagtcatatg agctttcccg attcgttgct caacccgctg caagacaacc  13080 

tcaccaagca aggcgcggtg gttcactcca tcggtgcgtg cggtgccggt gcggcggatt  13140 

gggttgtgcc gaaaaaagtc gaatgcggcg gcgaacgcac gcccaccggc aaggccgtga  13200 

tctatggcaa aaacgccatg agcaccacgc cgatccagga gctgatcgcc aaagacaaac  13260 

ccgacgtggt cgtgttgatc atcggcgaca ccatgggctc ctacaccaac ccggtgttcc  13320 

ctaaagcctg ggcctggaaa agcgtgacct cgctgaccaa agccatcacc gacaccggca  13380 

ccaagtgcgt gtgggtcggc ccgccatggg gcaaggtcgg ttcgcagtac aagaaagacg  13440 

acacccgcac caagctgatg tcgtcgttcc tcgccagtaa cgtggcgccg tgcacctaca  13500 

tcgactcgct gacgttctcg aagccaggcg agtggatcac caccgacggc cagcacttca  13560 

ccatcgacgg ctaccagaag tgggccaagg ccatcggtac agccttgggt gacctgccgc  13620 

catcggccta cggtaaagga aacaaataat gaaacggatg accctgggcc tggcgacctt  13680 

gctcgccagt gtcagcgcct actgtgccga catcccgctg tacccgaccg gcccggagca  13740 

agacgcagca ttcctgcgtt ttgccaacgg cacccctggc gaattgaagc tggtggcgga  13800 

cggttccaag gccagcctgg tgttgagcgg cgacaaagcc gtgtcggcgt tcctgccggt  13860 

tgtcggcggc gacaaaccga tcaaaggcgt attgagcagc ggcggcaaaa acgctgattt  13920 

ctcggtgaaa gtggcgccgg gcgaattcgc cacggtggtt gcactggtcg acgccaaggg  13980 

cgccacgcgc caattggtgg tgcgtgaagt gccggacgac ttcaatgcgc tcaaagcctc  14040 

actggcgttc atcaacgccg acgccacctg cgccgacgcc agcctggaag ccgtggcgca  14100 

aaaggccgag ctgttcaagc aggtcgctga aggtgccgtg cagcggcgca tgatcaaccc  14160 

ggtggaattg tcggtgcagc tcaaatgcgc cggctcgccg gtgggccagc cgttgacgtt  14220 

cactctcaag gcgggcgagc gctacagcgt attggccgtt ccttcagaca caggctcgaa  14280 

gttgctgttc gcctctgacg cactcgctaa ctgatcgggc ccgacaccac catggtcttt  14340 

gcctcactcg aattcctcac gctgttcctg ccggccttcc tgttgatcta tgccctggct  14400 

cgtccgagct ggcgcaacgt gatcctgttg atcggcagct ggttgttcta cggctggttg  14460 

agcccgttgt tcctgttcct gcacatggtg ttgaccgtgg tggcatgggt cggcggcttg  14520 

ctggtggacc gctcccgtga ggacggcaaa ggccgggtgc gcctgttgat cgcgttgatc  14580 

gtgttcaaca cggccgtgct gtgttggtac aagtacgcca acatcgtcgc cggcactgtg  14640 

agcgaggtga tcacctggta cggtgcgatg ccgttggact ggcagcgcgt ggccttgccg  14700 

gcgggcttgt cgttcatcgt cttgcaggcg atttcctacc tggtggatgt gcaccgtcat  14760 

acggtgccgg tggagcgcag cttcattaac tacgccacct atatctcgat gttcgggcac  14820 

tcgattgcgg gcccgatcat ccgttacgac tgggtccgcc gtgagctgaa ccagcggtat  14880 

ttcaactggg cgaatttctc cctgggcgca cggcgcttca tgatcggcat gggcatgaaa  14940 

gtgctggtgg ccgacacctt gtcgccgctg gtggacattg ccttccacct ggaaaacccc  15000 

agcctggtgg acgcctggat cggttgcttg gcgtattcgc tgcaactgtt tttcgacttc  15060 

gccggctaca gcgccatggc catcggcttg ggcttgatgc tgggcttcca cttcccggaa  15120 

aacttcaacc ggccgtacct gcagcagcat cagacttctg cggcgttgca cttgtcgtgt  15180 

cagctgctgc gcgactacct gtacatcgcg ctggcgggta accgtgacgg tgcctggcgc  15240 

acctaccgca acctgttcct gaccatggcg attgccgggt tgtggcacgg cggcgacagc  15300 

tggaactacc tgttgtgggg ttcggcccac ggcgtggcgc tgtgtgttga ccgtgcatgg  15360 

tcgcggtcga gcttgccgag cattccgccg gtgctgtcgc acgttctcac gctgctgttt  15420 

gtgtgcctgg cctggacgtt gttccgcgcg ccggacttcc attcggcact gaccatgtac  15480 

gccggccagt tcggtctgca cggcatggcc ttgggtgacg cgatggccgt ggccatgcgc  15540 

ccggcccatg gcatggcggc gttgctgggg ctggtgtgca tcatcgcgcc gatctggcag  15600 

gtgcgttgcg aacagcgctt cggcacccag ccgtggtttg tggtggcggc ctcgctgtgg  15660 

ccggtggccg ggtttgtgtt gtcgttcgcg ctgattgcca gccgcgacgc cgtaccgttt  15720 

ctgtactttc agttctaagg acgcccgacc atgcctgctc ctaccgcgcc gactccaccg  15780 

agcgacttgg ccgttcgcac cagccccttg gcggcctggg tgctggtgcc ctttttggcg  15840 

gcggggctgc tgtcctgcgt ttggctgatg gtgaaggggc cgatcagtta tgtgccggcc  15900 

aaggtcgaca gcgacatgtt gctgcacggt gacctgaccc accggtttgc caaggaattg  15960 

gccaaggcgc cgatggcgat ccaggccgcc aacctggagc gcggcggcag ctggctggcg  16020 

tttggcgaca ccgggccgcg cgtacgtccg ggttgccccg gctggctgtt tatcagcgat  16080 

gagctgcgca tcaaccggca tgccgaggcc aatgcccaga ccaaggccca ggcggtgatc  16140 

gacctgcaaa aacaactggg tcaaaaaggt atcgacctgc aagtggtggt ggtgccggac  16200 

aagagccgca tcgccgccgc ccaacgctgt ggcctgtacc gcccggcggt gctcgataac  16260 

cgggtgcggg attggaccgc catgttgcag gccgccggcg tttccgcgtt ggacctcacc  16320 

gaaacattga aaccgctggg cgccgaagcc tacctgcgca ccgacaccca ctggagcgaa  16380 

atcggctcca acgcgggggc caaggccgtg gcgcagcgta cccagcagcg cggcatcaag  16440 

gccacgccgg agcaaacctt cgacatcacc caggcgccgc tcgccgtgcg tccgggcgac  16500 

ttggtgcgcc tggccggtct cgactggttg ccgcccacgt tgcagccgcc aggggaatcg  16560 

gtcgcggcca gcacgaccca tgaaaccggt ggtgcgacca gtaatgccga cgatctgttt  16620 

ggcgatgccg gcctgcccaa tgtggcgctg atcggcacgt cgttctcgcg taactccaac  16680 

tttgtcggct tcctgcaaaa ggccctgaat gcgcctgtgg gcaatttcag caaggacggc  16740 

ggcgaattct ccggtgcggc caaggcgtac ttcgacagcc cggcgttcaa gcaaaccccg  16800 

cctaagttgt tgatttggga gattccagag cgcgatttgc aaacgccata cgatgtcatc  16860 

acgattggcc agtaatttga cgattgtccg tttgtctaaa aaataaacag cgcaagcatc  16920 

aaaataatga cactcttgtg agcttgaagc agactgcctg cacaagggcg acgacaaaaa  16980 

gacctgcggt aagcagccgg gtgagctggc cttttcacgt cgttaagctt cgccgtgatg  17040 

aggaatccgc tatgagcttt acagatggtc ttctcgttct gctggggaaa gacgtcagcc  17100 

gtgacgccca agggctggat gcgcggctgc atttttttgg cagcattccc gtggatgacg  17160 

gcaccccgtt tccccctcaa gcccccagtg cgcaggcgca atcccagtcc gtggataagg  17220 

gggtacgacg tccgtgcgtg gtggcgttgg tgtcggtcaa tggcggcgtg gggcgcagta  17280 

ccttggcgac ggccttgagc agcggtttgc agcgcctggg tgagtcggtg gtcgccgtgg  17340 

acctggaccc acagaacgcc ctgcggatgc acttcggtgt cagccccgcc tcgccgggta  17400 

tcggcccgac aagcctgcgc aatgcgcagt gggacaatat ccagcagcca ggctttgtcg  17460 

gcagtcgtgt gatcaccttt ggcgacaccg atatgcgcca gcaggacgac ctgcaacgtt  17520 

ggctcaaaca tgaaccggac tggttggcgc agcgcctgtc tgcattgggc ctgagcgccc  17580 

gtcataccgt gatcatcgat acccccgccg gcaataacgt ctacttccat caagccttga  17640 

gcgtggccga cgtggtgctg gtgattgccc aagccgacgc cgcctccctg ggtacgctgg  17700 

accaactcga cgggctgttg gcaccgcacc ttcagcgcga acgcccaccg catgtgcact  17760 

ttgtgatcaa ccaactggat gaggacaatg ccttcagcct ggatatggtg gaggcgttca  17820 

agcagcggct cggcaccaga gagcctctag aggtgcaccg cgatatggcg atcaagcgag  17880 

gcgctggcgt ttggtatcga cccattggat agccaggcat tgagcttggc gggggacgat  17940 

atcaccgagc tttgccggct gctgattgca cgtaaaaaac ggctttaaat tgcgctttat  18000 

tcgcttaacg ccagatgctt atgcacaatc ggttggaggg caatgtcccg aaacagccgt  18060 

tgtgtggagg cattctcatc acgccgcaac tgcctgtttt gagtgcgttt tattttgtga  18120 

acaaaaaatg accagcctgg cttttggcct gtaacgtaga tggctacggg ctctgtaaag  18180 

aattcatcaa cagagttatc cacaggatgt gccgcgacaa tttcgccctt taatcgcgct  18240 

cggcaagcac catcaagttg cgcggcgtca aggccggttc gcagaaggtg ccgacttcca  18300 

ccctgtagcc gttttcgccg agaaacagtg cacgatccag caccagccat aactccagcg  18360 

gccgccgaaa cagaccacgc accaattcca ggtttctgac ctctgccagg cgtcgccagc  18420 

catacgcctc taaaccggcc caatcctgtt cgtctgtgga taaccctttg agctttgcca  18480 

gcgcgtggca gtagtccgcg aagggcctgt ccagccagct tcgcgggcag cgacggcgtg  18540 

gacaggtact cgtcacagcc acgcaactgg cgttgcagtt gatcgaaacc caggcgtcgc  18600 

gccatggacg tgtcccgctg caaacgcacc cgtttgcccg cggtgacggt ttcgctcagc  18660 

ggcaggccga ggtcatcgat cgacagttgc aggggtgaag cacgaccggc gctggacagt  18720 

ggctggtagg cctgggcgtt tatgcggttg tagcagcagg gcgccagcgc caactgtttg  18780 

cagccggcgg cgctggccag gtgcagcagg cgtacatgca ggtcgccaca ggcatgcagg  18840 

gcgacgggcg tgtgttcggg gccgatagcc acagcggcca tcacgtcttg caggcaatgc  18900 

gtgacggcag gccgtgatgc tcgctcaagg cctggcctgc ggcgatcagc gccgggtcgt  18960 

attccaggca ggtcaatgct tgatctgcct gcaacaggcg acgacccaga tggccttttc  19020 

ccgcacacca gtccagccag tgcctgggct tttgcgcaaa ctgcagcgcg gcgccgaacg  19080 

cttcgatttg tgcccattta cggccaggta catcgacgct aaggcgatgg cgagcgggcg  19140 

ccaagggctg catcggtaac ttatccacag cactgagctg ccgggcttgc gcggcaagct  19200 

gcggaaacgg cgcgggtgcc gccaggtcgt ggggctgatt gtggctggct tcggcagcgg  19260 

ctaacgaacg tcggcgtagc cactgggcga gttcggggtg ctgggtttcc caaggtaaat  19320 

acaggtgcgt aaagggccgt ggtttccaca gcccttggtg gtcggtcagg aatgcatcca  19380 

gcgcctggaa gcgtgcctca acgtccttga caggcatcaa cgcgcagcca gcgttccagc  19440 

agtttgaagc cgcgtaccag caggtacgac atcagcaggt agaacacacc agcggtgaag  19500 

aacatatcca ctgtcaggta gttgcgcgca atgatcgtcc tggccatgcc ggtcagctca  19560 

agcagcgtta ccgtactcgc cagcgagctg gccttgagca tcaggatcac ttcgttgctg  19620 

taggccggca agccgatacg cgcggcgcgc ggcaggatga tgtagaacaa cgtcttgggc  19680 

ttggacatgc ccagggcgcg agcggcttcg atctcgcccg ggggaatagc ctggatcgca  19740 

ccgcgcagga tctcagcgat gtacgcggcg gtgtggaggg tcatggtggc cgtggcacac  19800 

cagaacggat cacgcaggta cggccacatg gcgctggtgc gcacggcatc gaactgggcc  19860 

aggccgtagt agaccaggaa cagttgcacc agcagcggtg tgccacggaa gaaaaagatg  19920 

taggcgtagg gcagcgaacg cacataccaa cgcttggacg cacgagcgac gcccagcgga  19980 

atcgcgagta tcaggccaag aatgacagcg atgcccacca gttccagggt caggatggcg  20040 

ccttggataa actttggcag ccacttgatg atgacttccc attccattac gtcgtcctca  20100 

cgaagccgcg ggcggcgcgt tgttccaaga agtgcatgcc gaccatcgac agcgaggtca  20160 

gggcccaggt agatgaaggc cgcgaccaga tagaaggtga agggttgccg gtaacggtga  20220 

ccgcatctgc gcgtgacgca tgattcttca ggccgatgac cgtaccagcg caggtcttat  20280 

aggatcatga cagtaccagc tgcagc                                       20306 

 
           
             2  
             344  
             PRT  
             Pseudomonas fluorescens  
           
            2 

Val Ser Arg Ala Asp Asp Ile Ser Lys Leu Phe Asn Lys Leu Gly Ala 
  1               5                  10                  15 

Asn Pro Ser Gly Tyr Arg Glu Ile Asp Phe Val His Glu Phe Ile Glu 
             20                  25                  30 

Asp Asp Val Glu Val Leu Glu Thr Pro Ala Val Val Arg Ala Leu Pro 
         35                  40                  45 

Val Ile Glu Ala Pro Ser Ala Pro Leu Leu Arg Leu Leu Glu Glu Leu 
     50                  55                  60 

Ser Gln Gly Glu Ala Asp His Leu Gln Pro Pro Glu Val Val Glu Gly 
 65                  70                  75                  80 

Arg Asp Gly Glu Val Tyr Ser Glu His Ser Ser Pro Asn Val Val Val 
                 85                  90                  95 

Val Val Ser Val Lys Gly Gly Val Gly Arg Ser Thr Leu Thr Ala Ala 
            100                 105                 110 

Ile Ala Ser Gly Leu Gln Arg Gln Gly Arg Pro Ala Leu Ala Leu Asp 
        115                 120                 125 

Leu Asp Pro Gln Asn Ala Leu Arg His His Leu Cys Leu Gly Leu Asp 
    130                 135                 140 

Met Pro Gly Val Gly Ala Thr Ser Leu Leu Asn Glu Ser Trp Glu Ala 
145                 150                 155                 160 

Leu Pro Glu Arg Gly Phe Ala Gly Cys Arg Leu Val Ala Phe Gly Ala 
                165                 170                 175 

Thr Asp His Glu Gln Gln Gln Ser Leu Asn Arg Trp Leu Gly Gln Asp 
            180                 185                 190 

Asp Glu Trp Leu Ser Lys Arg Leu Ala Gly Leu Lys Leu Asn Gly Gln 
        195                 200                 205 

Asp Thr Val Ile Ile Asp Val Pro Ala Gly Asn Thr Val Tyr Phe Ser 
    210                 215                 220 

Gln Ala Met Ser Val Ala Asp Ala Val Leu Val Val Val Gln Pro Asp 
225                 230                 235                 240 

Val Ala Ser Phe Ser Thr Leu Asp Gln Met Asp Ser Val Leu Lys Pro 
                245                 250                 255 

Ser Leu Asn Arg Lys Lys Thr Pro Arg Arg Phe Tyr Val Ile Asn Gln 
            260                 265                 270 

Leu Asp Gly Ala His Arg Phe Ser Leu Asp Met Ala Glu Val Phe Lys 
        275                 280                 285 

Thr Arg Leu Gly Ala Ala Leu Leu Gly Thr Val His Arg Asp Pro Ala 
    290                 295                 300 

Phe Ser Glu Ala Gln Ala Tyr Gly Arg Asp Pro Leu Asp Pro Thr Val 
305                 310                 315                 320 

Asn Ser Ile Gly Ser Gln Asp Ile His Ala Leu Cys Arg Ala Leu Leu 
                325                 330                 335 

Glu Arg Ile Asp Ser Asp Leu Pro 
            340 

 
           
             3  
             739  
             PRT  
             Pseudomonas fluorescens  
           
            3 

Met Thr Asp Thr Thr Ser Ser Thr Pro Phe Val Glu Gly Arg Ala Glu 
  1               5                  10                  15 

Gln Arg Leu Asn Gly Ala Ile Ala Arg Phe Asn Arg Trp Pro Ser Ala 
             20                  25                  30 

Pro Arg Thr Val Leu Val Val Ala Ser Cys Val Leu Gly Ala Met Leu 
         35                  40                  45 

Leu Leu Gly Ile Ile Ser Ala Pro Leu Asp Leu Tyr Ser Gln Cys Leu 
     50                  55                  60 

Phe Ala Ala Val Cys Phe Leu Ala Val Leu Val Leu Arg Lys Ile Pro 
 65                  70                  75                  80 

Gly Arg Leu Ala Ile Leu Ala Leu Val Val Leu Ser Leu Val Ala Ser 
                 85                  90                  95 

Leu Arg Tyr Met Phe Trp Arg Leu Thr Ser Thr Leu Gly Phe Glu Thr 
            100                 105                 110 

Trp Val Asp Met Phe Phe Gly Tyr Gly Leu Val Ala Ala Glu Phe Tyr 
        115                 120                 125 

Ala Leu Ile Val Leu Ile Phe Gly Tyr Val Gln Thr Ala Trp Pro Leu 
    130                 135                 140 

Arg Arg Thr Pro Val Trp Leu Lys Thr Glu Pro Glu Glu Trp Pro Thr 
145                 150                 155                 160 

Val Asp Val Phe Ile Pro Thr Tyr Asn Glu Ala Leu Ser Ile Val Lys 
                165                 170                 175 

Leu Thr Ile Phe Ala Ala Gln Ala Met Asp Trp Pro Lys Asp Lys Leu 
            180                 185                 190 

Arg Val His Val Leu Asp Asp Gly Arg Arg Asp Asp Phe Arg Glu Phe 
        195                 200                 205 

Cys Arg Lys Val Gly Val Asn Tyr Ile Arg Arg Asp Asn Asn Phe His 
    210                 215                 220 

Ala Lys Ala Gly Asn Leu Asn Glu Ala Leu Lys Val Thr Asp Gly Glu 
225                 230                 235                 240 

Tyr Ile Ala Leu Phe Asp Ala Asp His Val Pro Thr Arg Ser Phe Leu 
                245                 250                 255 

Gln Val Ser Leu Gly Trp Phe Leu Lys Asp Pro Lys Leu Ala Met Leu 
            260                 265                 270 

Gln Thr Pro His Phe Phe Phe Ser Pro Asp Pro Phe Glu Lys Asn Leu 
        275                 280                 285 

Asp Thr Phe Arg Ala Val Pro Asn Glu Gly Glu Leu Phe Tyr Gly Leu 
    290                 295                 300 

Val Gln Asp Gly Asn Asp Leu Trp Asn Ala Thr Phe Phe Cys Gly Ser 
305                 310                 315                 320 

Cys Ala Val Ile Arg Arg Glu Pro Leu Leu Glu Ile Gly Gly Val Ala 
                325                 330                 335 

Val Glu Thr Val Thr Glu Asp Ala His Thr Ala Leu Lys Leu Asn Arg 
            340                 345                 350 

Leu Gly Tyr Asn Thr Ala Tyr Leu Ala Ile Pro Gln Ala Ala Gly Leu 
        355                 360                 365 

Ala Thr Glu Ser Leu Ser Arg His Ile Asn Gln Arg Ile Arg Trp Ala 
    370                 375                 380 

Arg Gly Met Ala Gln Ile Phe Arg Thr Asp Asn Pro Leu Leu Gly Lys 
385                 390                 395                 400 

Gly Leu Lys Trp Gly Gln Arg Ile Cys Tyr Ala Asn Ala Met Gln His 
                405                 410                 415 

Phe Phe Tyr Gly Leu Pro Arg Leu Val Phe Leu Thr Ala Pro Leu Ala 
            420                 425                 430 

Tyr Leu Ile Phe Gly Ala Glu Ile Phe His Ala Ser Ala Leu Met Ile 
        435                 440                 445 

Val Ala Tyr Val Leu Pro His Leu Val His Ser Ser Leu Thr Asn Ser 
    450                 455                 460 

Arg Ile Gln Gly Arg Phe Arg His Ser Phe Trp Asn Glu Val Tyr Glu 
465                 470                 475                 480 

Thr Val Leu Ala Trp Tyr Ile Leu Pro Pro Val Leu Val Ala Leu Val 
                485                 490                 495 

Asn Pro Lys Ala Gly Gly Phe Asn Val Thr Asp Lys Gly Gly Ile Ile 
            500                 505                 510 

Asp Lys Gln Phe Phe Asp Trp Lys Leu Ala Arg Pro Tyr Leu Val Leu 
        515                 520                 525 

Leu Ala Val Asn Leu Ile Gly Leu Gly Phe Gly Ile His Gln Leu Ile 
    530                 535                 540 

Trp Gly Asp Ala Ser Thr Ala Val Thr Val Ala Ile Asn Leu Thr Trp 
545                 550                 555                 560 

Thr Leu Tyr Asn Leu Ile Ile Thr Ser Ala Ala Val Ala Val Ala Ser 
                565                 570                 575 

Glu Ala Arg Gln Val Arg Ser Glu Pro Arg Val Ser Ala Lys Leu Pro 
            580                 585                 590 

Val Ser Ile Ile Cys Ala Asp Gly Arg Val Leu Asp Gly Thr Thr Gln 
        595                 600                 605 

Asp Phe Ser Gln Asn Gly Phe Gly Leu Met Leu Ser Asp Gly His Ser 
    610                 615                 620 

Ile Thr Gln Gly Glu Arg Val Gln Leu Val Leu Ser Arg Asn Gly Gln 
625                 630                 635                 640 

Asp Ser Leu Lys Asp Ala Arg Val Val Phe Ser Lys Gly Ala Gln Ile 
                645                 650                 655 

Gly Ala Gln Phe Glu Ala Leu Ser Leu Arg Gln Gln Ser Glu Leu Val 
            660                 665                 670 

Arg Leu Thr Phe Ser Arg Ala Asp Thr Trp Ala Ala Ser Trp Gly Ala 
        675                 680                 685 

Gly Gln Pro Asp Thr Pro Leu Ala Ala Leu Arg Glu Val Gly Ser Ile 
    690                 695                 700 

Gly Ile Gly Gly Leu Phe Thr Leu Gly Arg Ala Thr Leu His Glu Leu 
705                 710                 715                 720 

Arg Leu Ala Leu Ser Arg Thr Pro Thr Lys Pro Leu Asp Thr Leu Met 
                725                 730                 735 

Asp Lys Pro 

 
           
             4  
             689  
             PRT  
             Pseudomonas fluorescens  
           
            4 

Val Trp Ser Arg Pro Gln Arg Asn Phe Asp Ile Arg Ala Asp Glu Val 
  1               5                  10                  15 

Val Lys Gly Ala Gln Leu Leu Leu Lys Tyr Ser Tyr Ser Pro Ala Leu 
             20                  25                  30 

Leu Ala Asp Leu Ser Gln Ile Asn Val Leu Val Asn Gly Glu Val Ala 
         35                  40                  45 

Ala Ser Leu Pro Leu Pro Lys Glu Gly Ala Gly Thr Pro Gln Glu Gln 
     50                  55                  60 

Leu Val Gln Ile Pro Ala His Leu Ile Thr Glu Phe Asn Arg Leu Ser 
 65                  70                  75                  80 

Leu Gln Phe Ile Gly His Tyr Thr Met Ser Cys Glu Asp Pro Leu His 
                 85                  90                  95 

Ser Ser Leu Trp Ala Lys Ile Ser Asn Ser Ser Glu Leu Lys Val Gln 
            100                 105                 110 

Val Glu Pro Ile Val Leu Lys Asp Asp Leu Ala Val Leu Pro Leu Pro 
        115                 120                 125 

Phe Phe Asp Lys Arg Asp Ala Arg Gln Val Ser Leu Pro Phe Val Phe 
    130                 135                 140 

Ala Thr Ala Pro Asp Ser Ala Ala Leu Glu Ala Ala Gly Ala Leu Ser 
145                 150                 155                 160 

Ser Trp Ile Gly Gly Leu Ala Ser Tyr Arg Gly Ala Thr Phe Pro Thr 
                165                 170                 175 

Thr Leu Gly Glu Leu Pro Ala Lys Gly Asn Ala Ile Val Leu Val Gln 
            180                 185                 190 

Thr Ala Asp Ala Met Asp Ile His Gly Val Ala Val Ala Lys Pro Ala 
        195                 200                 205 

Gly Pro Thr Leu Thr Leu Ile Ala Asn Pro Asn Asp Ala Asn Gly Lys 
    210                 215                 220 

Leu Leu Ile Val Thr Gly Arg Asp Gly Ala Glu Leu Lys Arg Ala Ala 
225                 230                 235                 240 

Thr Arg Trp Cys Trp Gln Pro Gly Ile Gly Arg Tyr Ser Val Val Ile 
                245                 250                 255 

Thr Lys Leu Asp Thr Leu Ala Pro Arg Arg Pro Tyr Asp Ala Pro Asn 
            260                 265                 270 

Trp Leu Pro Ser Asn Arg Pro Val Arg Leu Gly Glu Leu Ile Glu Gln 
        275                 280                 285 

Gln Lys Leu Ser Val Ser Gly Tyr Asn Pro Gly Ala Ile Ser Val Asp 
    290                 295                 300 

Met Arg Leu Pro Pro Asp Leu Phe Asn Trp Arg Glu Glu Gly Val Pro 
305                 310                 315                 320 

Leu Lys Leu Lys Tyr Arg Tyr Thr Pro Gln Gln Val Ser Thr Asn Ser 
                325                 330                 335 

Ser Leu Leu Ile Gly Leu Asn Asp Gln Phe Met Lys Ser Val Ala Leu 
            340                 345                 350 

Pro Ser Val Ser Asn Leu Gly Gly Gly Gln Thr Leu Leu Asp Gln Leu 
        355                 360                 365 

Lys Lys Asp Glu Ser Leu Pro Arg Glu Val Thr Thr Leu Leu Pro Ile 
    370                 375                 380 

Ser Ser Ala Ser Pro Lys Ser Lys Leu Gln Val Arg Phe Met Tyr Asp 
385                 390                 395                 400 

Tyr Ile Lys Glu Gly Glu Cys Arg Asp Ile Ile Val Asp Asn Met Arg 
                405                 410                 415 

Gly Ser Val Asp Pro Asp Ser Thr Leu Asp Val Thr Gly Tyr Gln His 
            420                 425                 430 

Tyr Ile Ala Met Pro Asn Leu Gly Val Phe Asn Asp Ser Gly Phe Pro 
        435                 440                 445 

Phe Thr Arg Leu Ala Asp Leu Ser Glu Ser Ala Val Val Met Pro Asp 
    450                 455                 460 

Asn Tyr Gly Thr Asp Glu Leu Thr Ala Tyr Leu Thr Val Leu Gly Arg 
465                 470                 475                 480 

Phe Gly Glu Ala Thr Gly Tyr Pro Ala Thr Ala Val Lys Val Val Gln 
                485                 490                 495 

Ala Lys Asp Val Gln Ser Val Ala Asp Lys Asp Leu Leu Val Leu Ala 
            500                 505                 510 

Thr Ala Ala Asn Gln Pro Leu Leu Lys Gln Trp Gln Gln Tyr Leu Pro 
        515                 520                 525 

Ala Thr Ser Asp Gly Glu Gln His Gln Phe Leu Leu Ser Asp Leu Pro 
    530                 535                 540 

Arg Tyr Val Arg Ser Trp Ile Ser Pro Asp Pro Ala Ala Asn Gln His 
545                 550                 555                 560 

Pro Ala Asn Thr Gly Ile Thr Phe Lys Gly Leu Ser Asn Ser Thr Trp 
                565                 570                 575 

Leu Ala Gly Phe Gln Ser Pro Leu Lys Ser Gly Arg Ser Val Val Leu 
            580                 585                 590 

Ile Ala Ser Asn Gln Pro Gln Gly Leu Leu Glu Ala Thr Asn Ala Leu 
        595                 600                 605 

Ile Gly Gly Asp Asp Tyr Lys Asp Ser Ile Gln Gly Ser Leu Ala Val 
    610                 615                 620 

Val Gln Gly Thr Gln Ile Ser Ser Leu Val Gly Asp Glu Gln Tyr Tyr 
625                 630                 635                 640 

Val Gly Lys Leu Asn Tyr Phe Lys Phe Met Gln Trp Gln Leu Ser Gln 
                645                 650                 655 

Asn Leu Gly Trp Met Leu Leu Ile Thr Phe Leu Gly Leu Ala Val Val 
            660                 665                 670 

Thr Ser Leu Ile Tyr Leu Ser Leu Arg Ala Arg Ala Lys Arg Arg Leu 
        675                 680                 685 

Ala 

 
           
             5  
             436  
             PRT  
             Pseudomonas fluorescens  
           
            5 

Val Ala Thr Val Ala Glu Pro Gly Leu Asp Ala Val Asp His Leu Pro 
  1               5                  10                  15 

Arg Pro Gly Arg Gly His Gln Pro Asp Leu Leu Val Ala Ala Cys Pro 
             20                  25                  30 

Cys Lys Thr Ala Val Gly Met Ser Pro Leu Lys Cys Met Ala Leu Ala 
         35                  40                  45 

Ala Leu Gly Ala Val Met Phe Val Gly Ser Ala Gln Ala Gln Thr Cys 
     50                  55                  60 

Asp Trp Pro Leu Trp Gln Asn Tyr Ala Lys Arg Phe Val Gln Asp Asp 
 65                  70                  75                  80 

Gly Arg Val Leu Asn Ser Ser Met Lys Pro Thr Glu Ser Ser Ser Glu 
                 85                  90                  95 

Gly Gln Ser Tyr Ala Met Phe Phe Ala Leu Val Gly Asn Asp Arg Ala 
            100                 105                 110 

Ser Phe Asp Lys Leu Trp Thr Trp Thr Lys Ala Asn Met Ser Gly Ala 
        115                 120                 125 

Asp Ile Gly Gln Asn Leu Pro Gly Trp Leu Trp Gly Lys Lys Ala Asp 
    130                 135                 140 

Asn Thr Trp Gly Val Ile Asp Pro Asn Ser Ala Ser Asp Ala Asp Leu 
145                 150                 155                 160 

Trp Met Ala Tyr Ala Leu Leu Glu Ala Ala Arg Val Trp Asn Ala Pro 
                165                 170                 175 

Gln Tyr Arg Ala Asp Ala Gln Leu Leu Leu Ala Asn Val Glu Arg Asn 
            180                 185                 190 

Leu Ile Val Arg Val Pro Gly Leu Gly Lys Met Leu Leu Pro Gly Pro 
        195                 200                 205 

Val Gly Tyr Val His Ala Gly Gly Leu Trp Arg Phe Asn Pro Ser Tyr 
    210                 215                 220 

Gln Val Leu Ala Gln Leu Arg Arg Phe His Lys Glu Arg Pro Asn Ala 
225                 230                 235                 240 

Gly Trp Asn Glu Val Ala Asp Ser Asn Ala Lys Met Leu Ala Asp Thr 
                245                 250                 255 

Ala Ser Asn Pro His Gly Leu Ala Ala Asn Trp Val Gly Tyr Arg Ala 
            260                 265                 270 

Thr Ser Ala Asn Thr Gly Leu Phe Val Val Asp Pro Phe Ser Asp Asp 
        275                 280                 285 

Leu Gly Ser Tyr Asp Ala Ile Arg Thr Tyr Met Trp Ala Gly Met Thr 
    290                 295                 300 

Ala Lys Gly Asp Pro Leu Ala Ala Pro Met Leu Lys Ser Leu Gly Gly 
305                 310                 315                 320 

Met Thr Arg Ala Thr Ala Ala Ser Ala Thr Gly Tyr Pro Pro Glu Lys 
                325                 330                 335 

Ile His Val Leu Thr Gly Glu Val Glu Lys Asn Asn Gly Tyr Thr Pro 
            340                 345                 350 

Met Gly Phe Ser Ala Ser Thr Val Ala Phe Phe Gln Ala Arg Gly Glu 
        355                 360                 365 

Thr Ala Leu Ala Gln Leu Gln Lys Ala Lys Val Asp Asp Ala Leu Ala 
    370                 375                 380 

Lys Ala Leu Ala Pro Ser Ala Pro Asp Thr Ala Gln Pro Ile Tyr Tyr 
385                 390                 395                 400 

Asp Tyr Met Leu Ser Leu Phe Ser Gln Gly Phe Ala Asp Gln Lys Tyr 
                405                 410                 415 

Arg Phe Glu Gln Asp Gly Thr Val Lys Leu Ser Trp Glu Ala Ala Cys 
            420                 425                 430 

Ala Val Thr Arg 
        435 

 
           
             6  
             1279  
             PRT  
             Pseudomonas fluorescens  
           
            6 

Met Arg Arg His Thr Leu Ala Ile Ala Ile Leu Ala Ala Leu Ala Ser 
  1               5                  10                  15 

Thr Ala Ser Val Ala Glu Thr Ser Asp Pro Gln Ser Leu Leu Ile Glu 
             20                  25                  30 

Gln Gly Tyr Tyr Trp Gln Ser Lys Lys Asn Pro Glu Arg Ala Leu Glu 
         35                  40                  45 

Thr Trp Gln Lys Leu Leu Arg Leu Ser Pro Asp Gln Pro Asp Ala Leu 
     50                  55                  60 

Tyr Gly Ile Gly Leu Ile Gln Val Gln Gln Asn His Pro Ala Glu Ala 
 65                  70                  75                  80 

Gln Lys Tyr Leu Ala Arg Leu Gln Ala Leu Ser Pro Val Pro Arg Gln 
                 85                  90                  95 

Ala Leu Gln Leu Glu Gln Asp Ile Thr Val Ala Val Pro Asp Asn Ala 
            100                 105                 110 

Lys Leu Leu Glu Gln Ala Arg Glu Leu Gly Glu Pro Glu Ala Glu Arg 
        115                 120                 125 

Glu Gln Ala Val Ala Leu Tyr Arg Gln Ile Phe Gln Gly Arg Gln Pro 
    130                 135                 140 

Gln Gly Leu Ile Ala Arg Glu Tyr Tyr Asn Thr Leu Gly Phe Thr Ala 
145                 150                 155                 160 

Lys Gly Ser Ser Glu Ala Ile Ala Gly Leu Gln Arg Leu Thr Arg Glu 
                165                 170                 175 

Arg Pro Asn Asp Pro Ile Val Ala Leu Phe Leu Ala Lys His Leu Ala 
            180                 185                 190 

Arg Asn Pro Ala Thr Arg Pro Asp Gly Ile Arg Ala Leu Ala Lys Leu 
        195                 200                 205 

Ala Ser Asn Asn Asp Val Gly Gly Asn Ala Asp Glu Thr Trp Arg Phe 
    210                 215                 220 

Ala Leu Val Trp Leu Gly Pro Pro Lys Pro Asp Gln Val Ser Leu Phe 
225                 230                 235                 240 

Gln Gln Phe Leu Thr Val His Pro Asp Asp Ser Glu Ile Arg Ala Leu 
                245                 250                 255 

Met Asn Lys Gly Ile Ala Gln Gly Lys Gly Gly Gly Thr Trp Gln Arg 
            260                 265                 270 

Asp Pro Gln Met Thr Lys Ala Phe Lys Ala Leu Asp Asp Gly Asp Leu 
        275                 280                 285 

Lys Thr Ala Glu Pro Leu Leu Ala Ala Arg Leu Ala Gln Lys Ser Asn 
    290                 295                 300 

Asp Val Asp Ala Leu Gly Gly Met Gly Val Leu Arg Gln Gln Gln Glu 
305                 310                 315                 320 

Arg Tyr Ser Glu Ala Glu Asn Tyr Leu Val Gln Ala Thr Arg Leu Pro 
                325                 330                 335 

Gly Gly Ala Ala Trp Gln Ser Ala Leu Asn Asp Val Arg Tyr Trp Asn 
            340                 345                 350 

Leu Ile Ser Gln Ser Arg Asp Ala Gln Arg Ala Gly Arg Ser Ala Gln 
        355                 360                 365 

Ala Arg Asp Leu Val Ala Gln Ala Glu Arg Leu Asn Pro Gly Gln Pro 
    370                 375                 380 

Gly Ala Ala Ile Ala Leu Ala Gly Phe Gln Ala Gln Asp Asn Gln Phe 
385                 390                 395                 400 

Asp Asp Ala Glu Ala Gly Tyr Arg Lys Val Leu Ala Arg His Pro Gly 
                405                 410                 415 

Asp Pro Asp Ala Leu Ser Gly Leu Ile Asn Val Leu Ser Gln Ser Gly 
            420                 425                 430 

Gln Pro Asp Glu Ala Leu Lys Leu Ile Asp Ser Val Ser Pro Ala Gln 
        435                 440                 445 

Arg Ala Lys Phe Ala Pro Ser Val Lys Ile Asn Ala Leu Arg Ala Thr 
    450                 455                 460 

Gln Val Gly Lys Leu Ala Glu Gln Arg Gly Asp Leu Lys Ala Ala Gln 
465                 470                 475                 480 

Ala Ala Tyr Arg Gln Ala Leu Asp Ala Asp Pro Glu Asn Pro Trp Thr 
                485                 490                 495 

Arg Phe Ala Leu Ala Arg Met Tyr Leu Arg Asp Gly Gln Ile Arg Asn 
            500                 505                 510 

Ala Arg Ala Leu Ile Asp Gly Leu Leu Lys Ser Gln Pro Asn Gln Pro 
        515                 520                 525 

Asp Ala Leu Tyr Thr Ser Thr Leu Leu Ser Ala Gln Leu Ser Glu Trp 
    530                 535                 540 

Lys Gln Ala Glu Ala Thr Leu Gly Arg Ile Pro Thr Ala Gln Arg Thr 
545                 550                 555                 560 

Ala Asp Met Asn Glu Leu Ala Thr Asp Ile Ala Leu His Gln Gln Thr 
                565                 570                 575 

Asp Ile Ala Ile Glu Thr Ala Arg Arg Gly Gln Arg Pro Glu Ala Leu 
            580                 585                 590 

Ala Leu Leu Gly Arg Ser Glu Pro Leu Thr Arg Asn Lys Pro Glu Arg 
        595                 600                 605 

Val Ala Val Leu Ala Ala Ala Tyr Val Glu Val Gly Ala Ala Gln Tyr 
    610                 615                 620 

Gly Leu Asp Met Met Gln Lys Val Val Glu Asn Asn Pro Asn Pro Thr 
625                 630                 635                 640 

Val Asp Gln Lys Leu Leu Tyr Ala Asn Val Leu Leu Lys Ala Asn Lys 
                645                 650                 655 

Tyr Ser Glu Ala Gly Glu Ile Leu Arg Glu Val Gln Gly Gln Pro Leu 
            660                 665                 670 

Thr Glu Thr Gly Arg Gln Arg Tyr Asp Asp Leu Ile Tyr Leu Tyr Arg 
        675                 680                 685 

Val Lys Gln Ala Asp Ala Leu Arg Glu Lys Asn Asp Leu Val Ala Ala 
    690                 695                 700 

Tyr Asp Met Leu Ser Pro Ala Leu Ala Gln Arg Pro Asn Asp Ala Leu 
705                 710                 715                 720 

Gly Val Gly Ala Leu Ala Arg Met Tyr Ala Ala Ser Gly Asn Gly Lys 
                725                 730                 735 

Lys Ala Met Glu Leu Tyr Ala Pro Leu Ile Gln Gln Asn Pro Asn Asn 
            740                 745                 750 

Ala Arg Leu Gln Leu Gly Leu Ala Asp Ile Ala Leu Lys Gly Asn Asp 
        755                 760                 765 

Arg Gly Leu Ala Gln Ser Ala Ser Asp Lys Ala Leu Ala Leu Glu Pro 
    770                 775                 780 

Gly Asn Pro Glu Ile Leu Thr Ser Ala Ala Arg Ile Tyr Gln Gly Leu 
785                 790                 795                 800 

Gly Lys Asn Ser Glu Ala Ala Glu Leu Leu Arg Lys Ala Leu Ala Ile 
                805                 810                 815 

Glu Asn Ala Met Lys Ala Lys Thr Gln Val Ala Gln Ala Ser Ala Pro 
            820                 825                 830 

Gly Thr Ser Tyr Asn Pro Phe Val Gly Leu Pro Gly Gln Arg Arg Gln 
        835                 840                 845 

Val Thr Asp Leu Thr Val Ala Gly Ala Val Pro Pro Pro Ile Asp Ala 
    850                 855                 860 

Pro Thr Lys Ser Val Thr Ser Asn Ala Phe Ala Ser Ala Thr Ser Asn 
865                 870                 875                 880 

Asp Leu Ser Asp Pro Phe Val Pro Pro Ser Ser Ile Ala Ser Ile Asp 
                885                 890                 895 

Ser Pro Glu Leu Ser Pro Ala Arg Arg Ala Leu Asp Thr Ile Leu Arg 
            900                 905                 910 

Asp Arg Thr Gly Tyr Val Val Gln Gly Leu Ser Val Arg Ser Asn Asn 
        915                 920                 925 

Gly Glu Lys Gly Leu Ser Lys Ile Thr Asp Val Glu Ala Pro Phe Glu 
    930                 935                 940 

Ala Arg Met Pro Val Gly Asp Asn Thr Val Ala Leu Arg Val Thr Pro 
945                 950                 955                 960 

Val His Leu Ser Ala Gly Ser Val Lys Ala Glu Ser Leu Ser Arg Phe 
                965                 970                 975 

Gly Lys Gly Gly Thr Glu Pro Ala Gly Ser Gln Ser Asp Ser Gly Val 
            980                 985                 990 

Gly Leu Ala Val Ala Phe Glu Asn Pro Asp Gln Gly Leu Lys Ala Asp 
        995                 1000                1005 

Val Gly Val Ser Pro Leu Gly Phe Leu Tyr Asn Thr Leu Val Gly Gly 
    1010                1015                1020 

Val Ser Val Ser Arg Pro Phe Glu Ala Asn Ser Asn Phe Arg Tyr Gly 
1025                1030                1035                1040 

Ala Asn Ile Ser Arg Arg Pro Val Thr Asp Ser Val Thr Ser Phe Ala 
                1045                1050                1055 

Gly Ser Glu Asp Gly Ala Gly Asn Lys Trp Gly Gly Val Thr Ala Asn 
            1060                1065                1070 

Gly Gly Arg Gly Glu Leu Ser Tyr Asp Asn Gln Lys Leu Gly Val Tyr 
        1075                1080                1085 

Gly Tyr Ala Ser Leu His Glu Leu Leu Gly Asn Asn Val Glu Asp Asn 
    1090                1095                1100 

Thr Arg Leu Glu Leu Gly Ser Gly Ile Tyr Trp Tyr Leu Arg Asn Asn 
1105                1110                1115                1120 

Pro Arg Asp Thr Leu Thr Leu Gly Ile Ser Gly Ser Ala Met Thr Phe 
                1125                1130                1135 

Lys Glu Asn Gln Asp Phe Tyr Thr Tyr Gly Asn Gly Gly Tyr Phe Ser 
            1140                1145                1150 

Pro Gln Arg Phe Phe Ser Leu Gly Val Pro Ile Arg Trp Ala Gln Ser 
        1155                1160                1165 

Phe Asp Arg Phe Ser Tyr Gln Val Lys Ser Ser Val Gly Leu Gln His 
    1170                1175                1180 

Ile Ala Gln Asp Gly Ala Asp Tyr Phe Pro Gly Asp Ser Thr Leu Gln 
1185                1190                1195                1200 

Ala Thr Lys Asn Asn Pro Lys Tyr Asp Ser Thr Ser Lys Thr Gly Val 
                1205                1210                1215 

Gly Tyr Ser Phe Asn Ala Ala Ala Glu Tyr Arg Leu Ser Ser Arg Phe 
            1220                1225                1230 

Tyr Leu Gly Gly Glu Ile Gly Leu Asp Asn Ala Gln Asp Tyr Arg Gln 
        1235                1240                1245 

Tyr Ala Gly Asn Ala Tyr Leu Arg Tyr Leu Phe Glu Asp Leu Ser Gly 
    1250                1255                1260 

Pro Met Pro Leu Pro Val Ser Pro Tyr Arg Ser Pro Tyr Ser Asn 
1265                1270                1275 

 
           
             7  
             221  
             PRT  
             Pseudomonas fluorescens  
           
            7 

Met Pro Val Ser Ala Ile Ala Gly Leu Thr Met Leu Val Leu Gly Glu 
  1               5                  10                  15 

Ser His Met Ser Phe Pro Asp Ser Leu Leu Asn Pro Leu Gln Asp Asn 
             20                  25                  30 

Leu Thr Lys Gln Gly Ala Val Val His Ser Ile Gly Ala Cys Gly Ala 
         35                  40                  45 

Gly Ala Ala Asp Trp Val Val Pro Lys Lys Val Glu Cys Gly Gly Glu 
     50                  55                  60 

Arg Thr Pro Thr Gly Lys Ala Val Ile Tyr Gly Lys Asn Ala Met Ser 
 65                  70                  75                  80 

Thr Thr Pro Ile Gln Glu Leu Ile Ala Lys Asp Lys Pro Asp Val Val 
                 85                  90                  95 

Val Leu Ile Ile Gly Asp Thr Met Gly Ser Tyr Thr Asn Pro Val Phe 
            100                 105                 110 

Pro Lys Ala Trp Ala Trp Lys Ser Val Thr Ser Leu Thr Lys Ala Ile 
        115                 120                 125 

Thr Asp Thr Gly Thr Lys Cys Val Trp Val Gly Pro Pro Trp Gly Lys 
    130                 135                 140 

Val Gly Ser Gln Tyr Lys Lys Asp Asp Thr Arg Thr Lys Leu Met Ser 
145                 150                 155                 160 

Ser Phe Leu Ala Ser Asn Val Ala Pro Cys Thr Tyr Ile Asp Ser Leu 
                165                 170                 175 

Thr Phe Ser Lys Pro Gly Glu Trp Ile Thr Thr Asp Gly Gln His Phe 
            180                 185                 190 

Thr Ile Asp Gly Tyr Gln Lys Trp Ala Lys Ala Ile Gly Thr Ala Leu 
        195                 200                 205 

Gly Asp Leu Pro Pro Ser Ala Tyr Gly Lys Gly Asn Lys 
    210                 215                 220 

 
           
             8  
             221  
             PRT  
             Pseudomonas fluorescens  
           
            8 

Met Lys Arg Met Thr Leu Gly Leu Ala Thr Leu Leu Ala Ser Val Ser 
  1               5                  10                  15 

Ala Tyr Cys Ala Asp Ile Pro Leu Tyr Pro Thr Gly Pro Glu Gln Asp 
             20                  25                  30 

Ala Ala Phe Leu Arg Phe Ala Asn Gly Thr Pro Gly Glu Leu Lys Leu 
         35                  40                  45 

Val Ala Asp Gly Ser Lys Ala Ser Leu Val Leu Ser Gly Asp Lys Ala 
     50                  55                  60 

Val Ser Ala Phe Leu Pro Val Val Gly Gly Asp Lys Pro Ile Lys Gly 
 65                  70                  75                  80 

Val Leu Ser Ser Gly Gly Lys Asn Ala Asp Phe Ser Val Lys Val Ala 
                 85                  90                  95 

Pro Gly Glu Phe Ala Thr Val Val Ala Leu Val Asp Ala Lys Gly Ala 
            100                 105                 110 

Thr Arg Gln Leu Val Val Arg Glu Val Pro Asp Asp Phe Asn Ala Leu 
        115                 120                 125 

Lys Ala Ser Leu Ala Phe Ile Asn Ala Asp Ala Thr Cys Ala Asp Ala 
    130                 135                 140 

Ser Leu Glu Ala Val Ala Gln Lys Ala Glu Leu Phe Lys Gln Val Ala 
145                 150                 155                 160 

Glu Gly Ala Val Gln Arg Arg Met Ile Asn Pro Val Glu Leu Ser Val 
                165                 170                 175 

Gln Leu Lys Cys Ala Gly Ser Pro Val Gly Gln Pro Leu Thr Phe Thr 
            180                 185                 190 

Leu Lys Ala Gly Glu Arg Tyr Ser Val Leu Ala Val Pro Ser Asp Thr 
        195                 200                 205 

Gly Ser Lys Leu Leu Phe Ala Ser Asp Ala Leu Ala Asn 
    210                 215                 220 

 
           
             9  
             468  
             PRT  
             Pseudomonas fluorescens  
           
            9 

Met Val Phe Ala Ser Leu Glu Phe Leu Thr Leu Phe Leu Pro Ala Phe 
  1               5                  10                  15 

Leu Leu Ile Tyr Ala Leu Ala Arg Pro Ser Trp Arg Asn Val Ile Leu 
             20                  25                  30 

Leu Ile Gly Ser Trp Leu Phe Tyr Gly Trp Leu Ser Pro Leu Phe Leu 
         35                  40                  45 

Phe Leu His Met Val Leu Thr Val Val Ala Trp Val Gly Gly Leu Leu 
     50                  55                  60 

Val Asp Arg Ser Arg Glu Asp Gly Lys Gly Arg Val Arg Leu Leu Ile 
 65                  70                  75                  80 

Ala Leu Ile Val Phe Asn Thr Ala Val Leu Cys Trp Tyr Lys Tyr Ala 
                 85                  90                  95 

Asn Ile Val Ala Gly Thr Val Ser Glu Val Ile Thr Trp Tyr Gly Ala 
            100                 105                 110 

Met Pro Leu Asp Trp Gln Arg Val Ala Leu Pro Ala Gly Leu Ser Phe 
        115                 120                 125 

Ile Val Leu Gln Ala Ile Ser Tyr Leu Val Asp Val His Arg His Thr 
    130                 135                 140 

Val Pro Val Glu Arg Ser Phe Ile Asn Tyr Ala Thr Tyr Ile Ser Met 
145                 150                 155                 160 

Phe Gly His Ser Ile Ala Gly Pro Ile Ile Arg Tyr Asp Trp Val Arg 
                165                 170                 175 

Arg Glu Leu Asn Gln Arg Tyr Phe Asn Trp Ala Asn Phe Ser Leu Gly 
            180                 185                 190 

Ala Arg Arg Phe Met Ile Gly Met Gly Met Lys Val Leu Val Ala Asp 
        195                 200                 205 

Thr Leu Ser Pro Leu Val Asp Ile Ala Phe His Leu Glu Asn Pro Ser 
    210                 215                 220 

Leu Val Asp Ala Trp Ile Gly Cys Leu Ala Tyr Ser Leu Gln Leu Phe 
225                 230                 235                 240 

Phe Asp Phe Ala Gly Tyr Ser Ala Met Ala Ile Gly Leu Gly Leu Met 
                245                 250                 255 

Leu Gly Phe His Phe Pro Glu Asn Phe Asn Arg Pro Tyr Leu Gln Gln 
            260                 265                 270 

His Gln Thr Ser Ala Ala Leu His Leu Ser Cys Gln Leu Leu Arg Asp 
        275                 280                 285 

Tyr Leu Tyr Ile Ala Leu Ala Gly Asn Arg Asp Gly Ala Trp Arg Thr 
    290                 295                 300 

Tyr Arg Asn Leu Phe Leu Thr Met Ala Ile Ala Gly Leu Trp His Gly 
305                 310                 315                 320 

Gly Asp Ser Trp Asn Tyr Leu Leu Trp Gly Ser Ala His Gly Val Ala 
                325                 330                 335 

Leu Cys Val Asp Arg Ala Trp Ser Arg Ser Ser Leu Pro Ser Ile Pro 
            340                 345                 350 

Pro Val Leu Ser His Val Leu Thr Leu Leu Phe Val Cys Leu Ala Trp 
        355                 360                 365 

Thr Leu Phe Arg Ala Pro Asp Phe His Ser Ala Leu Thr Met Tyr Ala 
    370                 375                 380 

Gly Gln Phe Gly Leu His Gly Met Ala Leu Gly Asp Ala Met Ala Val 
385                 390                 395                 400 

Ala Met Arg Pro Ala His Gly Met Ala Ala Leu Leu Gly Leu Val Cys 
                405                 410                 415 

Ile Ile Ala Pro Ile Trp Gln Val Arg Cys Glu Gln Arg Phe Gly Thr 
            420                 425                 430 

Gln Pro Trp Phe Val Val Ala Ala Ser Leu Trp Pro Val Ala Gly Phe 
        435                 440                 445 

Val Leu Ser Phe Ala Leu Ile Ala Ser Arg Asp Ala Val Pro Phe Leu 
    450                 455                 460 

Tyr Phe Gln Phe 
465 

 
           
             10  
             374  
             PRT  
             Pseudomonas fluorescens  
           
            10 

Met Pro Ala Pro Thr Ala Pro Thr Pro Pro Ser Asp Leu Ala Val Arg 
  1               5                  10                  15 

Thr Ser Pro Leu Ala Ala Trp Val Leu Val Pro Phe Leu Ala Ala Gly 
             20                  25                  30 

Leu Leu Ser Cys Val Trp Leu Met Val Lys Gly Pro Ile Ser Tyr Val 
         35                  40                  45 

Pro Ala Lys Val Asp Ser Asp Met Leu Leu His Gly Asp Leu Thr His 
     50                  55                  60 

Arg Phe Ala Lys Glu Leu Ala Lys Ala Pro Met Ala Ile Gln Ala Ala 
 65                  70                  75                  80 

Asn Leu Glu Arg Gly Gly Ser Trp Leu Ala Phe Gly Asp Thr Gly Pro 
                 85                  90                  95 

Arg Val Arg Pro Gly Cys Pro Gly Trp Leu Phe Ile Ser Asp Glu Leu 
            100                 105                 110 

Arg Ile Asn Arg His Ala Glu Ala Asn Ala Gln Thr Lys Ala Gln Ala 
        115                 120                 125 

Val Ile Asp Leu Gln Lys Gln Leu Gly Gln Lys Gly Ile Asp Leu Gln 
    130                 135                 140 

Val Val Val Val Pro Asp Lys Ser Arg Ile Ala Ala Ala Gln Arg Cys 
145                 150                 155                 160 

Gly Leu Tyr Arg Pro Ala Val Leu Asp Asn Arg Val Arg Asp Trp Thr 
                165                 170                 175 

Ala Met Leu Gln Ala Ala Gly Val Ser Ala Leu Asp Leu Thr Glu Thr 
            180                 185                 190 

Leu Lys Pro Leu Gly Ala Glu Ala Tyr Leu Arg Thr Asp Thr His Trp 
        195                 200                 205 

Ser Glu Ile Gly Ser Asn Ala Gly Ala Lys Ala Val Ala Gln Arg Thr 
    210                 215                 220 

Gln Gln Arg Gly Ile Lys Ala Thr Pro Glu Gln Thr Phe Asp Ile Thr 
225                 230                 235                 240 

Gln Ala Pro Leu Ala Val Arg Pro Gly Asp Leu Val Arg Leu Ala Gly 
                245                 250                 255 

Leu Asp Trp Leu Pro Pro Thr Leu Gln Pro Pro Gly Glu Ser Val Ala 
            260                 265                 270 

Ala Ser Thr Thr His Glu Thr Gly Gly Ala Thr Ser Asn Ala Asp Asp 
        275                 280                 285 

Leu Phe Gly Asp Ala Gly Leu Pro Asn Val Ala Leu Ile Gly Thr Ser 
    290                 295                 300 

Phe Ser Arg Asn Ser Asn Phe Val Gly Phe Leu Gln Lys Ala Leu Asn 
305                 310                 315                 320 

Ala Pro Val Gly Asn Phe Ser Lys Asp Gly Gly Glu Phe Ser Gly Ala 
                325                 330                 335 

Ala Lys Ala Tyr Phe Asp Ser Pro Ala Phe Lys Gln Thr Pro Pro Lys 
            340                 345                 350 

Leu Leu Ile Trp Glu Ile Pro Glu Arg Asp Leu Gln Thr Pro Tyr Asp 
        355                 360                 365 

Val Ile Thr Ile Gly Gln 
    370 

 
           
             11  
             324  
             PRT  
             Pseudomonas fluorescens  
           
            11 

Val Ser Leu Lys Gln Thr Ala Cys Thr Arg Ala Thr Thr Lys Arg Pro 
  1               5                  10                  15 

Ala Val Ser Ser Arg Val Ser Trp Pro Phe His Val Val Lys Leu Arg 
             20                  25                  30 

Arg Asp Glu Glu Ser Ala Met Ser Phe Thr Asp Gly Leu Leu Val Leu 
         35                  40                  45 

Leu Gly Lys Asp Val Ser Arg Asp Ala Gln Gly Leu Asp Ala Arg Leu 
     50                  55                  60 

His Phe Phe Gly Ser Ile Pro Val Asp Asp Gly Thr Pro Phe Pro Pro 
 65                  70                  75                  80 

Gln Ala Pro Ser Ala Gln Ala Gln Ser Gln Ser Val Asp Lys Gly Val 
                 85                  90                  95 

Arg Arg Pro Cys Val Val Ala Leu Val Ser Val Asn Gly Gly Val Gly 
            100                 105                 110 

Arg Ser Thr Leu Ala Thr Ala Leu Ser Ser Gly Leu Gln Arg Leu Gly 
        115                 120                 125 

Glu Ser Val Val Ala Val Asp Leu Asp Pro Gln Asn Ala Leu Arg Met 
    130                 135                 140 

His Phe Gly Val Ser Pro Ala Ser Pro Gly Ile Gly Pro Thr Ser Leu 
145                 150                 155                 160 

Arg Asn Ala Gln Trp Asp Asn Ile Gln Gln Pro Gly Phe Val Gly Ser 
                165                 170                 175 

Arg Val Ile Thr Phe Gly Asp Thr Asp Met Arg Gln Gln Asp Asp Leu 
            180                 185                 190 

Gln Arg Trp Leu Lys His Glu Pro Asp Trp Leu Ala Gln Arg Leu Ser 
        195                 200                 205 

Ala Leu Gly Leu Ser Ala Arg His Thr Val Ile Ile Asp Thr Pro Ala 
    210                 215                 220 

Gly Asn Asn Val Tyr Phe His Gln Ala Leu Ser Val Ala Asp Val Val 
225                 230                 235                 240 

Leu Val Ile Ala Gln Ala Asp Ala Ala Ser Leu Gly Thr Leu Asp Gln 
                245                 250                 255 

Leu Asp Gly Leu Leu Ala Pro His Leu Gln Arg Glu Arg Pro Pro His 
            260                 265                 270 

Val His Phe Val Ile Asn Gln Leu Asp Glu Asp Asn Ala Phe Ser Leu 
        275                 280                 285 

Asp Met Val Glu Ala Phe Lys Gln Arg Leu Gly Thr Arg Glu Pro Leu 
    290                 295                 300 

Glu Val His Arg Asp Met Ala Ile Lys Arg Gly Ala Gly Val Trp Tyr 
305                 310                 315                 320 

Arg Pro Ile Gly 

 
           
             12  
             333  
             PRT  
             Pseudomonas fluorescens  
           
            12 

Met Asn Asp Leu Gln Ile Asp Asp Ile Lys Thr Asp Glu Asn Ala Ala 
  1               5                  10                  15 

Met Val Leu Leu Val Asp Asp Gln Ala Met Ile Gly Glu Ala Val Arg 
             20                  25                  30 

Arg Gly Leu Ala His Glu Glu Asn Ile Asp Phe His Phe Cys Ala Asp 
         35                  40                  45 

Pro His Gln Ala Ile Ala Gln Ala Ile Arg Ile Lys Pro Thr Val Ile 
     50                  55                  60 

Leu Gln Asp Leu Val Met Pro Gly Leu Asp Gly Leu Thr Leu Val Arg 
 65                  70                  75                  80 

Glu Tyr Arg Asn His Pro Ala Thr Gln Asn Ile Pro Ile Ile Val Leu 
                 85                  90                  95 

Ser Thr Lys Glu Asp Pro Leu Ile Lys Ser Ala Ala Phe Ser Ala Gly 
            100                 105                 110 

Ala Asn Asp Tyr Leu Val Lys Leu Pro Asp Asn Ile Glu Leu Val Ala 
        115                 120                 125 

Arg Ile Arg Tyr His Ser Arg Ser Tyr Met Thr Leu Leu Gln Arg Asp 
    130                 135                 140 

Ala Ala Tyr Arg Ala Leu Arg Val Ser Gln Gln Gln Leu Leu Asp Thr 
145                 150                 155                 160 

Asn Leu Val Leu Gln Arg Leu Met Asn Ser Asp Gly Leu Thr Gly Leu 
                165                 170                 175 

Ser Asn Arg Arg His Phe Asp Glu Tyr Leu Glu Leu Glu Trp Arg Arg 
            180                 185                 190 

Ala Met Arg Asp Gln Thr Gln Leu Ser Leu Leu Met Ile Asp Val Asp 
        195                 200                 205 

Phe Phe Lys Thr Tyr Asn Asp Ser Phe Gly His Val Glu Gly Asp Glu 
    210                 215                 220 

Ala Leu Arg Lys Val Ala Ala Thr Ile Arg Glu Ala Ser Ser Arg Pro 
225                 230                 235                 240 

Ser Asp Leu Pro Ala Arg Tyr Gly Gly Glu Glu Phe Ala Leu Val Leu 
                245                 250                 255 

Pro Asn Thr Ser Pro Gly Gly Ala Arg Leu Val Ala Glu Lys Leu Arg 
            260                 265                 270 

Met Ala Val Ala Ala Leu Lys Ile Pro His Ile Ala Pro Thr Glu Gly 
        275                 280                 285 

Ser Ser Leu Thr Ile Ser Ile Gly Leu Ser Thr Met Thr Pro Gln Gln 
    290                 295                 300 

Gly Thr Asp Cys Arg Gln Val Ile Val Ala Ala Asp Lys Gly Leu Tyr 
305                 310                 315                 320 

Thr Ala Lys His Asn Gly Arg Asn Gln Val Gly Ile Glu 
                325                 330 

 
           
             13  
             1002  
             DNA  
             Pseudomonas fluorescens  
           
            13 

atgaatgatt tacagatcga cgacatcaag accgacgaaa acgccgccat ggtgttgctg     60 

gtcgacgacc aggccatgat cggtgaagcc gtgcggcgtg gcctggccca tgaagaaaat    120 

atcgacttcc acttctgcgc cgacccacac caggcgattg cccaggcgat ccgtatcaag    180 

ccgaccgtta tcctgcagga tctggtgatg ccaggtctgg acggcctgac actggtgcgc    240 

gagtaccgca accacccggc cacgcagaac atcccgatca tcgtgctttc caccaaggaa    300 

gacccgctga tcaagagcgc ggcgttttcg gccggggcca acgattattt ggtcaagctg    360 

ccggacaaca tcgagctggt ggcgcgcatc cgctatcact cgcgctccta catgaccctg    420 

ttgcaacggg atgcggctta tcgcgcgttg cgggtcagcc agcagcagct gttggacacc    480 

aacctggtgc tgcaacggct gatgaactcc gatggcctca cggggctgtc caaccgtcgc    540 

catttcgacg agtacctgga actggaatgg cgccgtgcca tgcgtgatca gactcagctg    600 

tcgttgttga tgattgatgt ggatttcttc aagacctaca acgatagctt tgggcatgtc    660 

gaaggtgacg aggctttgcg caaggttgcg gcgaccattc gtgaggccag cagtcggcct    720 

tcggatttgc cggcgcgcta tgggggggag gagtttgccc tggtgctgcc taatacctcg    780 

ccgggcgggg cgcggttggt ggctgagaag ctgcggatgg cggttgccgc gctgaaaatt    840 

ccgcacattg cgccgactga ggggtcgagt ctgaccatca gtattgggct gtcgaccatg    900 

acgccgcagc aggggacgga ttgtcggcag gtgatagtgg cggcggataa ggggttgtat    960 

acggctaagc ataatgggcg caatcaggtg gggattgagt ag                      1002 

 
           
             14  
             333  
             PRT  
             Pseudomonas fluorescens  
           
            14 

Met Asn Asp Leu Gln Ile Asp Asp Ile Lys Thr Asp Glu Asn Ala Ala 
  1               5                  10                  15 

Met Val Leu Leu Val Asp Asp Gln Ala Met Ile Gly Glu Ala Val Arg 
             20                  25                  30 

Arg Gly Leu Ala His Glu Glu Asn Ile Asp Phe His Phe Cys Ala Asp 
         35                  40                  45 

Pro His Gln Ala Ile Ala Gln Ala Ile Arg Ile Lys Pro Thr Val Ile 
     50                  55                  60 

Leu Gln Asp Leu Val Met Pro Gly Leu Asp Gly Leu Thr Leu Val Arg 
 65                  70                  75                  80 

Glu Tyr Arg Asn His Pro Ala Thr Gln Asn Ile Pro Ile Ile Val Leu 
                 85                  90                  95 

Ser Thr Lys Glu Asp Pro Leu Ile Lys Ser Ala Ala Phe Ser Ala Gly 
            100                 105                 110 

Ala Asn Asp Tyr Leu Val Lys Leu Pro Asp Asn Ile Glu Leu Val Ala 
        115                 120                 125 

Arg Ile Arg Tyr His Ser Arg Ser Tyr Met Thr Leu Leu Gln Arg Asp 
    130                 135                 140 

Ala Ala Tyr Arg Ala Leu Arg Val Ser Gln Gln Gln Leu Leu Asp Thr 
145                 150                 155                 160 

Asn Leu Val Leu Gln Arg Leu Met Asn Ser Asp Gly Leu Thr Gly Leu 
                165                 170                 175 

Ser Asn Arg Arg His Phe Asp Glu Tyr Leu Glu Leu Glu Trp Arg Arg 
            180                 185                 190 

Ala Met Arg Asp Gln Thr Gln Leu Ser Leu Leu Met Ile Asp Val Asp 
        195                 200                 205 

Phe Phe Lys Thr Tyr Asn Asp Ser Phe Gly His Val Glu Gly Asp Glu 
    210                 215                 220 

Ala Leu Arg Lys Val Ala Ala Thr Ile Arg Glu Ala Ser Ser Arg Pro 
225                 230                 235                 240 

Ser Asp Leu Pro Ala Arg Tyr Gly Gly Glu Glu Ser Ala Leu Val Leu 
                245                 250                 255 

Pro Asn Thr Ser Pro Gly Gly Ala Arg Leu Val Ala Glu Lys Leu Arg 
            260                 265                 270 

Met Ala Val Ala Ala Leu Lys Ile Pro His Ile Ala Pro Thr Glu Gly 
        275                 280                 285 

Ser Ser Leu Thr Ile Ser Ile Gly Leu Ser Thr Met Thr Pro Gln Gln 
    290                 295                 300 

Gly Thr Asp Cys Arg Gln Val Ile Val Ala Ala Asp Lys Gly Leu Tyr 
305                 310                 315                 320 

Thr Ala Lys His Asn Gly Arg Asn Gln Val Gly Ile Glu 
                325                 330 

 
           
             15  
             1002  
             DNA  
             Pseudomonas fluorescens  
           
            15 

atgaatgatt tacagatcga cgacatcaag accgacgaaa acgccgccat ggtgttgctg     60 

gtcgacgacc aggccatgat cggtgaagcc gtgcggcgtg gcctggccca tgaagaaaat    120 

atcgacttcc acttctgcgc cgacccacac caggcgattg cccaggcgat ccgtatcaag    180 

ccgaccgtta tcctgcagga tctggtgatg ccaggtctgg acggcctgac actggtgcgc    240 

gagtaccgca accacccggc cacgcagaac atcccgatca tcgtgctttc caccaaggaa    300 

gacccgctga tcaagagcgc ggcgttttcg gccggggcca acgattattt ggtcaagctg    360 

ccggacaaca tcgagctggt ggcgcgcatc cgctatcact cgcgctccta catgaccctg    420 

ttgcaacggg atgcggctta tcgcgcgttg cgggtcagcc agcagcagct gttggacacc    480 

aacctggtgc tgcaacggct gatgaactcc gatggcctca cggggctgtc caaccgtcgc    540 

catttcgacg agtacctgga actggaatgg cgccgtgcca tgcgtgatca gactcagctg    600 

tcgttgttga tgattgatgt ggatttcttc aagacctaca acgatagctt tgggcatgtc    660 

gaaggtgacg aggctttgcg caaggttgcg gcgaccattc gtgaggccag cagtcggcct    720 

tcggatttgc cggcgcgcta tgggggggag gagtctgccc tggtgctgcc taatacctcg    780 

ccgggcgggg cgcggttggt ggctgagaag ctgcggatgg cggttgccgc gctgaaaatt    840 

ccgcacattg cgccgactga ggggtcgagt ctgaccatca gtattgggct gtcgaccatg    900 

acgccgcagc aggggacgga ttgtcggcag gtgatagtgg cggcggataa ggggttgtat    960 

acggctaagc ataatgggcg caatcaggtg gggattgagt ag                      1002 

 
           
             16  
             333  
             PRT  
             Pseudomonas fluorescens  
           
            16 

Met Asn Asp Leu Gln Ile Asp Asp Ile Lys Thr Asp Glu Asn Ala Ala 
  1               5                  10                  15 

Met Val Leu Leu Val Asp Asp Gln Ala Met Ile Gly Glu Ala Val Arg 
             20                  25                  30 

Arg Gly Leu Ala His Glu Glu Asn Ile Asp Phe His Phe Cys Ala Asp 
         35                  40                  45 

Pro His Gln Ala Ile Ala Gln Ala Ile Arg Ile Lys Pro Thr Val Ile 
     50                  55                  60 

Leu Gln Asp Leu Val Met Pro Gly Leu Asp Gly Leu Thr Leu Val Arg 
 65                  70                  75                  80 

Glu Tyr Arg Asn His Pro Ala Thr Gln Asn Ile Pro Ile Ile Val Leu 
                 85                  90                  95 

Ser Thr Lys Glu Asp Pro Leu Ile Lys Ser Ala Ala Phe Ser Ala Gly 
            100                 105                 110 

Ala Asn Asp Tyr Leu Val Lys Leu Pro Asp Asn Ile Glu Leu Val Ala 
        115                 120                 125 

Arg Ile Arg Tyr His Ser Arg Ser Tyr Met Thr Leu Leu Gln Arg Asp 
    130                 135                 140 

Ala Ala Tyr Arg Ala Leu Arg Val Ser Gln Gln Gln Leu Leu Asp Thr 
145                 150                 155                 160 

Asn Leu Val Leu Gln Arg Leu Met Asn Ser Asp Gly Leu Thr Gly Leu 
                165                 170                 175 

Ser Asn Arg Arg His Phe Asp Glu Tyr Leu Glu Leu Glu Trp Arg Arg 
            180                 185                 190 

Ala Met Arg Asp Gln Thr Gln Leu Ser Leu Leu Met Ile Asp Val Asp 
        195                 200                 205 

Phe Phe Lys Thr Tyr Asn Asp Ser Phe Gly His Val Glu Gly Asp Glu 
    210                 215                 220 

Ala Leu Arg Lys Val Ala Ala Thr Ile Arg Glu Ala Ser Ser Arg Pro 
225                 230                 235                 240 

Ser Asp Leu Pro Ala Arg Tyr Gly Gly Glu Glu Phe Ala Leu Val Leu 
                245                 250                 255 

Pro Asn Thr Ser Pro Gly Gly Ala Arg Leu Val Ala Glu Lys Leu Arg 
            260                 265                 270 

Met Ala Val Ala Ala Leu Lys Ile Pro His Ile Ala Pro Thr Glu Gly 
        275                 280                 285 

Ser Ser Leu Thr Ile Ser Ile Gly Leu Ser Thr Met Thr Pro Gln Gln 
    290                 295                 300 

Gly Thr Asp Cys Arg Gln Val Ile Val Ala Ala Asp Lys Gly Leu Tyr 
305                 310                 315                 320 

Thr Ala Lys His Asn Gly Arg Asn Gln Val Gly Ile Glu 
                325                 330 

 
           
             17  
             1002  
             DNA  
             Pseudomonas fluorescens  
           
            17 

atgaatgatt tacagatcga cgacatcaag accgacgaaa acgccgccat ggtgttgctg     60 

gtcgacgacc aggccatgat cggtgaagcc gtgcggcgtg gcctggccca tgaagaaaat    120 

atcgacttcc acttctgcgc cgacccacac caggcgattg cccaggcgat ccgtatcaag    180 

ccgaccgtta tcctgcagga tctggtgatg ccaggtctgg acggcctgac actggtgcgc    240 

gagtaccgca accacccggc cacgcagaac atcccgatca tcgtgctttc caccaaggaa    300 

gacccgctga tcaagagcgc ggcgttttcg gccggggcca acgattattt ggtcaagctg    360 

ccggacaaca tcgagctggt ggcgcgcatc cgctatcact cgcgctccta catgaccctg    420 

ttgcaacggg atgcggctta tcgcgcgttg cgggtcagcc agcagcagct gttggacacc    480 

aacctggtgc tgcaacggct gatgaactcc gatggcctca cggggctgtc caaccgtcgc    540 

catttcgacg agtacctgga actggaatgg cgccgtgcca tgcgtgatca gactcagctg    600 

tcgttgttga tgattgatgt ggatttcttc aagacctaca acgatagctt tgggcatgtc    660 

gaaggtgacg aggctttgcg caaggttgcg gcgaccattc gtgaggccag cagtcggcct    720 

tcggatttgc cggcgcgcta tgggggggag gagtttgccc tggtgctgcc taatacctcg    780 

ccgggcgggg cgcggttggt ggctgagaag ctgcggatgg cggttgccgc gctgaaaatt    840 

ccgcacattg cgccgactga ggggtcgagt ctgaccatca gtattaggct gtcgaccatg    900 

acgccgcagc aggggacgga ttgtcggcag gtgatagtgg cggcggataa ggggttgtat    960 

acggctaagc ataatgggcg caatcaggtg gggattgagt ag                      1002 

 
           
             18  
             333  
             PRT  
             Pseudomonas fluorescens  
           
            18 

Met Asn Asp Leu Gln Ile Asp Asp Ile Lys Thr Asp Glu Asn Ala Ala 
  1               5                  10                  15 

Met Val Leu Leu Val Asp Asp Gln Ala Met Ile Gly Glu Ala Val Arg 
             20                  25                  30 

Arg Gly Leu Ala His Glu Glu Asn Ile Asp Phe His Phe Cys Ala Asp 
         35                  40                  45 

Pro His Gln Ala Ile Ala Gln Ala Ile Arg Ile Lys Pro Thr Val Ile 
     50                  55                  60 

Leu Gln Asp Leu Val Met Pro Gly Leu Asp Gly Leu Thr Leu Val Arg 
 65                  70                  75                  80 

Glu Tyr Arg Asn His Pro Ala Thr Gln Asn Ile Pro Ile Ile Val Leu 
                 85                  90                  95 

Ser Thr Lys Glu Asp Pro Leu Ile Lys Ser Ala Ala Phe Ser Ala Gly 
            100                 105                 110 

Ala Asn Asp Tyr Leu Val Lys Leu Pro Asp Asn Ile Glu Leu Val Ala 
        115                 120                 125 

Arg Ile Arg Tyr His Ser Arg Ser Tyr Met Thr Leu Leu Gln Arg Asp 
    130                 135                 140 

Ala Ala Tyr Arg Ala Leu Arg Val Ser Gln Gln Gln Leu Leu Asp Thr 
145                 150                 155                 160 

Asn Leu Val Leu Gln Arg Leu Met Asn Ser Asp Gly Leu Thr Gly Leu 
                165                 170                 175 

Ser Asn Arg Arg His Phe Asp Glu Tyr Leu Glu Leu Glu Trp Arg Arg 
            180                 185                 190 

Ala Met Arg Asp Gln Thr Gln Leu Ser Leu Leu Met Ile Asp Val Asp 
        195                 200                 205 

Phe Phe Lys Thr Tyr Asn Asp Ser Phe Gly His Val Glu Gly Asp Glu 
    210                 215                 220 

Ala Leu Arg Lys Val Ala Ala Thr Ile Arg Glu Ala Ser Ser Arg Pro 
225                 230                 235                 240 

Ser Asp Leu Pro Ala Arg Tyr Gly Gly Glu Glu Phe Ala Leu Val Leu 
                245                 250                 255 

Pro Asn Thr Ser Pro Gly Gly Ala Arg Leu Val Ala Glu Lys Leu Arg 
            260                 265                 270 

Met Ala Val Ser Ala Leu Lys Ile Pro His Ile Ala Pro Thr Glu Gly 
        275                 280                 285 

Ser Ser Leu Thr Ile Ser Ile Gly Leu Ser Thr Met Thr Pro Gln Gln 
    290                 295                 300 

Gly Thr Asp Cys Arg Gln Val Ile Val Ala Ala Asp Lys Gly Leu Cys 
305                 310                 315                 320 

Thr Ala Lys His Asn Gly Arg Asn Gln Val Gly Ile Glu 
                325                 330 

 
           
             19  
             1002  
             DNA  
             Pseudomonas fluorescens  
           
            19 

atgaatgatt tacagatcga cgacatcaag accgacgaaa acgccgccat ggtgttgctg     60 

gtcgacgacc aggccatgat cggtgaagcc gtgcggcgtg gcctggccca tgaagaaaat    120 

atcgacttcc acttctgcgc cgacccacac caggcgattg cccaggcgat ccgtatcaag    180 

ccgaccgtta tcctgcagga tctggtgatg ccaggtctgg acggcctgac actggtgcgc    240 

gagtaccgca accacccggc cacgcagaac atcccgatca tcgtgctttc caccaaggaa    300 

gacccgctga tcaagagcgc ggcgttttcg gccggggcca acgattattt ggtcaagctg    360 

ccggacaaca tcgagctggt ggcgcgcatc cgctatcact cgcgctccta catgaccctg    420 

ttgcaacggg atgcggctta tcgcgcgttg cgggtcagcc agcagcagct gttggacacc    480 

aacctggtgc tgcaacggct gatgaactcc gatggcctca cggggctgtc caaccgtcgc    540 

catttcgacg agtacctgga actggaatgg cgccgtgcca tgcgtgatca gactcagctg    600 

tcgttgttga tgattgatgt ggatttcttc aagacctaca acgatagctt tgggcatgtc    660 

gaaggtgacg aggctttgcg caaggttgcg gcgaccattc gtgaggccag cagtcggcct    720 

tcggatttgc cggcgcgcta tgggggggag gagtttgccc tggtgctgcc taatacctcg    780 

ccgggcgggg cgcggttggt ggctgagaag ctgcggatgg cggttaccgc gctgaaaatt    840 

ccgcacattg cgccgactga ggggtcgagt ctgaccatca gtattgggct gtcgaccatg    900 

acgccgcagc aggggacgga ttgtcggcag gtgatagtgg cggcggataa ggggttgtgt    960 

acggctaagc ataatgggcg caatcaggtg gggattgagt ag                      1002 

 
           
             20  
             333  
             PRT  
             Pseudomonas fluorescens  
           
            20 

Met Asn Asp Leu Gln Ile Asp Asp Ile Lys Thr Asp Glu Asn Ala Ala 
  1               5                  10                  15 

Met Val Leu Leu Val Asp Asp Gln Ala Met Ile Gly Glu Ala Val Arg 
             20                  25                  30 

Arg Gly Leu Ala His Glu Glu Asn Ile Asp Phe His Phe Cys Ala Asp 
         35                  40                  45 

Pro His Gln Ala Ile Ala Gln Ala Ile Arg Ile Lys Pro Thr Val Ile 
     50                  55                  60 

Leu Gln Asp Leu Val Met Pro Gly Leu Asp Gly Leu Thr Leu Val Arg 
 65                  70                  75                  80 

Glu Tyr Arg Asn His Pro Ala Thr Gln Asn Ile Pro Ile Ile Val Leu 
                 85                  90                  95 

Ser Thr Lys Glu Asp Pro Leu Ile Lys Ser Ala Ala Phe Ser Ala Gly 
            100                 105                 110 

Ala Asn Asp Tyr Leu Val Lys Leu Pro Asp Asn Ile Glu Leu Val Ala 
        115                 120                 125 

Arg Ile Arg Tyr His Ser Arg Ser Tyr Met Thr Leu Leu Gln Arg Asp 
    130                 135                 140 

Ala Ala Tyr Arg Ala Leu Arg Val Ser Gln Gln Gln Leu Leu Gly Thr 
145                 150                 155                 160 

Asn Leu Val Leu Gln Arg Leu Met Asn Ser Asp Gly Leu Thr Gly Leu 
                165                 170                 175 

Ser Asn Arg Arg His Phe Asp Glu Tyr Leu Glu Leu Glu Trp Arg Arg 
            180                 185                 190 

Ala Met Arg Asp Gln Thr Gln Leu Ser Leu Leu Met Ile Asp Val Asp 
        195                 200                 205 

Phe Phe Lys Thr Tyr Asn Asp Ser Phe Gly His Val Glu Gly Asp Glu 
    210                 215                 220 

Ala Leu Arg Lys Val Ala Ala Thr Ile Arg Glu Ala Ser Ser Arg Pro 
225                 230                 235                 240 

Ser Asp Leu Pro Ala Arg Tyr Gly Gly Glu Glu Phe Ala Leu Val Leu 
                245                 250                 255 

Pro Asn Thr Ser Pro Gly Gly Ala Arg Leu Val Ala Glu Lys Leu Arg 
            260                 265                 270 

Met Ala Val Ala Ala Leu Lys Ile Pro His Ile Ala Pro Thr Glu Gly 
        275                 280                 285 

Ser Ser Leu Thr Ile Ser Ile Gly Leu Ser Thr Met Thr Pro Gln Gln 
    290                 295                 300 

Gly Thr Asp Cys Arg Gln Val Ile Val Ala Ala Asp Lys Gly Leu Tyr 
305                 310                 315                 320 

Thr Ala Lys His Asn Gly Arg Asn Gln Val Gly Ile Glu 
                325                 330 

 
           
             21  
             1002  
             DNA  
             Pseudomonas fluorescens  
           
            21 

atgaatgatt tacagatcga cgacatcaag accgacgaaa acgccgccat ggtgttgctg     60 

gtcgacgacc aggccatgat cggtgaagcc gtgcggcgtg gcctggccca tgaagaaaat    120 

atcgacttcc acttctgcgc cgacccacac caggcgattg cccaggcgat ccgtatcaag    180 

ccgaccgtta tcctgcagga tctggtgatg ccaggtctgg acggcctgac actggtgcgc    240 

gagtaccgca accacccggc cacgcagaac atcccgatca tcgtgctttc caccaaggaa    300 

gacccgctga tcaagagcgc ggcgttttcg gccggggcca acgattattt ggtcaagctg    360 

ccggacaaca tcgagctggt ggcgcgcatc cgctatcact cgcgctccta catgaccctg    420 

ttgcaacggg atgcggctta tcgcgcgttg cgggtcagcc agcagcagct gttgggcacc    480 

aacctggtgc tgcaacggct gatgaactcc gatggcctca cggggctgtc caaccgtcgc    540 

catttcgacg agtacctgga actggaatgg cgccgtgcca tgcgtgatca gactcagctg    600 

tcgttgttga tgattgatgt ggatttcttc aagacctaca acgatagctt tgggcatgtc    660 

gaaggtgacg aggctttgcg caaggttgcg gcgaccattc gtgaggccag cagtcggcct    720 

tcggatttgc cggcgcgcta tgggggggag gagtttgccc tggtgctgcc taatacctcg    780 

ccgggcgggg cgcggttggt ggctgagaag ctgcggatgg cggttgccgc gctgaaaatt    840 

ccgcacattg cgccgactga ggggtcgagt ctgaccatca gtattgggct gtcgaccatg    900 

acgccgcagc aggggacgga ttgtcggcag gtgatagtgg cggcggataa ggggttgtat    960 

acggctaagc ataatgggcg caatcaggtg gggattgagt ag                      1002 

 
           
             22  
             333  
             PRT  
             Pseudomonas fluorescens  
           
            22 

Met Asn Asp Leu Gln Ile Asp Asp Ile Lys Thr Asp Glu Asn Ala Ala 
  1               5                  10                  15 

Met Val Leu Leu Val Asp Asp Gln Ala Met Ile Gly Glu Ala Val Arg 
             20                  25                  30 

Arg Gly Leu Ala His Glu Glu Asn Ile Asp Phe His Phe Cys Ala Asp 
         35                  40                  45 

Pro His Gln Ala Ile Ala Gln Ala Ile Arg Ile Lys Pro Thr Val Ile 
     50                  55                  60 

Leu Gln Asp Leu Val Met Pro Gly Leu Asp Gly Leu Thr Leu Val Arg 
 65                  70                  75                  80 

Glu Tyr Arg Asn His Pro Ala Thr Gln Asn Ile Pro Ile Ile Val Leu 
                 85                  90                  95 

Ser Thr Lys Glu Asp Pro Leu Ile Lys Ser Ala Ala Phe Ser Ala Gly 
            100                 105                 110 

Ala Asn Asp Tyr Leu Val Lys Leu Pro Asp Asn Ile Glu Leu Val Ala 
        115                 120                 125 

Cys Ile Arg Tyr His Ser Arg Ser Tyr Met Thr Leu Leu Gln Arg Asp 
    130                 135                 140 

Ala Ala Tyr Arg Ala Leu Arg Val Ser Gln Gln Gln Leu Leu Asp Thr 
145                 150                 155                 160 

Asn Leu Val Leu Gln Arg Leu Met Asn Ser Asp Gly Leu Thr Gly Leu 
                165                 170                 175 

Ser Asn Arg Arg His Phe Asp Glu Tyr Leu Glu Leu Glu Trp Arg Arg 
            180                 185                 190 

Ala Met Arg Asp Gln Thr Gln Leu Ser Leu Leu Met Ile Asp Val Asp 
        195                 200                 205 

Phe Phe Lys Thr Tyr Asn Asp Ser Phe Gly His Val Glu Gly Asp Glu 
    210                 215                 220 

Ala Leu Arg Lys Val Ala Ala Thr Ile Arg Glu Ala Ser Ser Arg Pro 
225                 230                 235                 240 

Ser Asp Leu Pro Ala Arg Tyr Gly Gly Glu Glu Phe Ala Leu Val Leu 
                245                 250                 255 

Pro Asn Thr Ser Pro Gly Gly Ala Arg Leu Val Ala Glu Lys Leu Arg 
            260                 265                 270 

Met Ala Val Ala Ala Leu Lys Ile Pro His Ile Ala Pro Thr Glu Gly 
        275                 280                 285 

Ser Ser Leu Thr Ile Ser Ile Gly Leu Ser Thr Met Thr Pro Gln Gln 
    290                 295                 300 

Gly Thr Asp Cys Arg Gln Val Ile Val Ala Ala Asp Lys Gly Leu Tyr 
305                 310                 315                 320 

Thr Ala Lys His Asn Gly Arg Asn Gln Val Gly Ile Glu 
                325                 330 

 
           
             23  
             1002  
             DNA  
             Pseudomonas fluorescens  
           
            23 

atgaatgatt tacagatcga cgacatcaag accgacgaaa acgccgccat ggtgttgctg     60 

gtcgacgacc aggccatgat cggtgaagcc gtgcggcgtg gcctggccca tgaagaaaat    120 

atcgacttcc acttctgcgc cgacccacac caggcgattg cccaggcgat ccgtatcaag    180 

ccgaccgtta tcctgcagga tctggtgatg ccaggtctgg acggcctgac actggtgcgc    240 

gagtaccgca accacccggc cacgcagaac atcccgatca tcgtgctttc caccaaggaa    300 

gacccgctga tcaagagcgc ggcgttttcg gccggggcca acgattattt ggtcaagctg    360 

ccggacaaca tcgagctggt ggcgtgcatc cgctatcact cgcgctccta catgaccctg    420 

ttgcaacggg atgcggctta tcgcgcgttg cgggtcagcc agcagcagct gttggacacc    480 

aacctggtgc tgcaacggct gatgaactcc gatggcctca cggggctgtc caaccgtcgc    540 

catttcgacg agtacctgga actggaatgg cgccgtgcca tgcgtgatca gactcagctg    600 

tcgttgttga tgattgatgt ggatttcttc aagacctaca acgatagctt tgggcatgtc    660 

gaaggtgacg aggctttgcg caaggttgcg gcgaccattc gtgaggccag cagtcggcct    720 

tcggatttgc cggcgcgcta tgggggggag gagtttgccc tggtgctgcc taatacctcg    780 

ccgggcgggg cgcggttggt ggctgagaag ctgcggatgg cggttgccgc gctgaaaatt    840 

ccgcacattg cgccgactga ggggtcgagt ctgaccatca gtattgggct gtcgaccatg    900 

acgccgcagc aggggacgga ttgtcggcag gtgatagtgg cggcggataa ggggttgtat    960 

acggctaagc ataatgggcg caatcaggtg gggattgagt ag                      1002 

 
           
             24  
             1136  
             DNA  
             Pseudomonas fluorescens  
             
               misc_feature  
               (174)..(174)  
               n = a, c, g or t/u  
             
           
            24 

ggtacctggc tcagggtggc tcggaagccg agttgtatcg tcacttcatc gccgtatcca     60 

gcaacaacgc ggcagccgtg gccttcggta tccgcgaaga aaacatcttc ccgatgtggg    120 

actgggtcgg cggccgttac tcactgtggt cggccatcgg cctgtctctc catnggcttg    180 

cccatcgccc tggcgatcgg catgtccaac ttcaaggaac tgctgtccag gtgcctacac    240 

catggaccag catttccaga acgccccatt cgaagccaac aagccggtgc tgctgggctt    300 

gctgggcgtg tggtacggca acttctgggg tgcgcagagc cacgcgatcc tgccgtacga    360 

ccactacctg cgtaacatca ccaagcactt gcaacagctg gacatggaat ccaacggcaa    420 

gagcgttcgc caggacggta cgcccgtcgc taccgatacc ggcccggtga tctggggtgg    480 

cgtgggttgc aacggccagc acgcttacca ccagttgctg catcagggga ccaactgatc    540 

ccggccgact tgcatcgtgc cgatcgtcag cttcaacccg gtgtccgacc accatcagtg    600 

gctgtacgcc aactgcctgt cccagagcca ggcgctgatn ctcggcaaga cccgcgtcga    660 

agccgaagcc gagctgcgcg acaaaggcat cccggaagac gaagtgcaga agctggcacc    720 

gcacaaggtg atcccgggca accgtccgag caacaccctc gtggtcgagc gcatcagccc    780 

gcgtcgctgg ggcactggtc gccatgtatg agcacaaagt gttcgtgcag agcgtgatct    840 

ggggcatcaa cgccttcgac caatggggcg tggaactggg caaagagctg ggcaaaggcg    900 

tctacaaccg cctgaccggc gccgaagaaa cctcggccga ggacgcttcg acccagggcc    960 

tgatcaacta cttccgcggt cgtcaccgcg gctgatacag gtattgcagg gccaacgtcc   1020 

gtttggaggt tggccttgaa cccttccttt gcatggtgca tctntatgac ttgtggctaa   1080 

aacaagaata aggatcgctc atgttcgata tcagcacgtt tcccaccgcc gatgcc       1136 

 
           
             25  
             703  
             DNA  
             Pseudomonas fluorescens  
             
               misc_feature  
               (5)..( 5)  
               n = a, c, g or t/u  
             
           
            25 

gtcangtcat ggaatctcag tgaaacagac gagctgtcgt gtgtagatgt cngaacacta     60 

cgcattctac ggtcttacgg ncgcnnttgt gggggttggt gcacagaaag tcggccaatt    120 

tagcacattg gcgccttgcc caaaatccct gtcgttgtta gagtttgccg cactttttta    180 

cccaggttgc ctagggtccc tttcccatgt tcaagaaact ggtggatgtt ttccagcgat    240 

ctttccattg acctggcact gccaacaccc ttatttacgt gcgagcgcgg tatcgtcctg    300 

aatgagccat cggttgtggc cattcggacc atggtaatca gaaaagtgtc gttncgtcgg    360 

caccgaggcc aagcgcatgc tcggccgtac accaggcaat attgctgcca ttcgtccgat    420 

gaaagacggc gtgatcgccg acttcagtgt ctgcgagaag atgctgcagt acttcatcaa    480 

caaggttcac gaaaacagtt tcctgcagcc cagccctcgt gtgctgatct gcgttccatg    540 

caaatccacc caggttgagc gtcgtgccat ccgtgaatcg gcccttggtg ccggtgctcg    600 

cgaagtgttc ctgatcgaag agccgatggc tgctgcgatc ggtgccggcc tgccggtaga    660 

agaagcccgc ggttcgatgg tggtggattc ggtggtggta ccc                      703 

 
           
             26  
             14000  
             DNA  
             Escherichia coli  
           
            26 

aacggaaagt caaaaagtga gcaaattccc gtctcgccgt aaacaataaa ccggcgaaat     60 

gctcacggtt agaattgtct catgaacggt acggttattt catagggatc aagcaaaatg    120 

aataacaatg aaccagatac tctgcctgat cccgcgatag gctatatctt ccagaatgat    180 

attgtggcgt taaagcaggc attttcactg cctgatattg attatgccga tatttcccaa    240 

cgcgaacagt tggccgcggc attaaaacgc tggccgttgc tggcagagtt tgcgcaacaa    300 

aagtagggga ttggtgaatg gccgtactgg gatagcaggg ggtgcgggga ggcgtgggga    360 

caacaaccat caccgccgca ttagcctggt cattacaaat gttgggagaa aatgtcctgg    420 

tggtcgatgc ctgcccggac aacttgttgc gcctgtcatt taatgttgat tttacccacc    480 

gtcagggctg ggccagagcg atgctggatg gccaggactg gcgtgacgct gggttgcgct    540 

acacctcgca gctcgatttg ctgccttttg gtcagttatc cattgaagaa caagaaaatc    600 

cacagcactg gcaaacccgg ctgagcgata tttgctccgg cttacagcaa ctaaaagcca    660 

gcgggcgtta ccagtggatt ttaatcgact taccgcgtga tgcctcgcag ataacccacc    720 

agctgctgag tttgtgcgat cactcgctgg caatcgtcaa tgtggatgcc aactgccata    780 

tccgactgca tcagcaagcg ctgccggatg gcgcacatat tttgattaat gacttccgta    840 

ttggcagtca ggttcaggac gatatttacc agctttggtt gcaaagccag cgccgattac    900 

tgccgatgct cattcatcgt gatgaagcga tggctgaatg cctggcggct aagcaaccag    960 

taggtgaata tcgcagtgat gcgctggcgg ctgaagagat actgacgctg gcgaactggt   1020 

gcctgttgaa ctactccggg ctgaaaacgc cagtcgggag tgcatcatga gtatcctgac   1080 

ccggtggttg cttatcccgc cggtcaacgc gcggcttatc gggcgttatc gcgattatcg   1140 

tcgtcacggt gcgtcggctt tcagcgcgac gctcggctgt ttctggatga tcctggcctg   1200 

gatttttatt ccgctggagc acccgcgctg gcagcgtatt cgcgcagaac ataaaaacct   1260 

gtatccgcat atcaacgcct cgcgtccgcg tccgctggac ccggtccgtt atctcattca   1320 

aacatgctgg ttattgatcg gtgcatcgcg caaagaaacg ccgaaaccgc gcaggcgggc   1380 

attttcaggt ctgcaaaata ttcgtggacg ttaccatcaa tggatgaacg agctgcctga   1440 

gcgcgttagc cataaaacac agcatctgga tgagaaaaaa gagctcggtc atttgagtgc   1500 

cggggcgcgg cggttgatcc tcggtatcat cgtcaccttc tcgctgattc tggcgttaat   1560 

ctgcgttact cagccgttta acccgctggc gcagtttatc ttcctgatgc tgctgtgggg   1620 

ggtagcgctg atcgtacggc ggatgccggg gcgcttctcg gcgctaatgt tgattgtgct   1680 

gtcgctgacc gtttcttgcc gttatatctg gtggcgttac acctctacgc tgaactggga   1740 

cgatccggtc agcctggtgt gcgggcttat tctgctcttc gctgaaacgt acgcgtggat   1800 

tgtgctggtg ctcggctact tccaggtagt atggccgctg aatcgtcagc cggtgccatt   1860 

gccgaaagat atgtcgctgt ggccgtcggt ggatatcttt gtcccgactt acaacgaaga   1920 

tctcaacgtg gtgaaaaata ccatttacgc ctcgctgggt atcgactggc cgaaagataa   1980 

gctgaatatc tggatccttg atgacggcgg cagggaagag tttcgccagt ttgcgcaaaa   2040 

cgtgggggtg aaatatatcg cccgcaccac tcatgaacat gcgaaagcag gcaacatcaa   2100 

caatgcgctg aaatatgcca aaggcgagtt cgtgtcgatt ttcgactgcg accacgtacc   2160 

aacgcgatcg ttcttgcaaa tgaccatggg ctggttcctg aaagaaaaac agctggcgat   2220 

gatgcagacg ccgcaccact tcttctcacc ggacccgttt gaacgcaacc tggggcgttt   2280 

ccgtaaaacg ccgaacgaag gcacgctgtt ctatggtctg gtgcaggatg gcaacgatat   2340 

gtgggacgcc actttcttct gcggttcctg tgcggtgatt cgtcgtaagc cgctggatga   2400 

aattggcggc attgctgtcg aaaccgtgac tgaagatgcg catacttctc tgcggttgca   2460 

ccgtcgtggc tatacctccg cgtatatgcg tattccgcag gcggcggggc tggcgaccga   2520 

aagtctgtcg gcgcatatcg gtcagcgtat tcgctgggcg cgcgggatgg tacaaatctt   2580 

ccgtctcgat aacccgctca ccggtaaagg gctgaagttt gctcagcggc tatgttacgt   2640 

caacgccatg ttccacttct tgtcgggcat tccacggctg atcttcctga ctgcgccgct   2700 

ggcgttcctg ctgcttcatg cctacatcat ctatgcgcca gcgttgatga tcgccctatt   2760 

cgtgctgccg catatgatcc atgccagcct gaccaactcc aagatccagg gcaaatatcg   2820 

ccactctttc tggagtgaaa tctacgaaac ggtgctggcg tggtatatcg caccaccgac   2880 

gctggtggcg ctgattaacc cgcacaaagg caaatttaac gtcaccgcca aaggtggact   2940 

ggtggaagaa gagtacgtcg actgggtgat ctcgcggccc tacatcttcc ttgtcctgct   3000 

caacctggtg ggcgttgcgg taggcatctg gcgctacttc tatggcccgc caaccgagat   3060 

gctcaccgtg gtcgtcagta tggtgtgggt gttctacaac ctgattgttc ttggcggcgc   3120 

agttgcggta tcggtagaaa gcaaacaggt acgccgatcg caccgcgtgg agatgacgat   3180 

gcccgcggca attgcccgcg aagatggtca cctcttctcg tgtaccgttc aggatttctc   3240 

cgacggtggt ttggggatca agatcaacgg tcaggcgcag attctggaag ggcagaaagt   3300 

gaatctgttg cttaaacgcg gtcagcagga atacgtcttc ccgacccagg tggcgcgcgt   3360 

gatgggtaat gaagttgggc tgaaattaat gccgctcacc acccagcaac atatcgattt   3420 

tgtgcagtgt acgtttgccc gtgcggatac atgggcgctc tggcaggaca gctacccgga   3480 

agataagccg ctggaaagtc tgctggatat tctgaagctc ggcttccgtg gctaccgcca   3540 

tctggcggag tttgcgcctt cttcggtgaa gggcatattc cgtgtgctga cttctctggt   3600 

ttcctgggtt gtatcgttta ttccgcgccg cccggagcgg agcgaaacgg cacaaccatc   3660 

ggatcaggct ttggctcaac aatgatgata acgcgatgaa aagaaaacta ttctggattt   3720 

gtgcagtggc tatggggatg agtgcgttcc cctctttcat gacgcaggcg acgccagcaa   3780 

cgcaaccact gatcaatgct gagccagctg tagccgccca gacggaacaa aatccgcagg   3840 

tggggcaagt gatgccgggc gtgcagggcg ctgatgcgcc agtcgtggcg cagaacggtc   3900 

cttcgcgtga tgtgaagctg acctttgcgc aaattgcacc gccgccgggc agcatggtgc   3960 

tacgtggcat taacccgaac ggcagcattg agtttggtat gcgcagcgat gaagtggtga   4020 

cgaaggcgat gctcaacctc gaatacaccc catcgccatc gttactgcct gtccagtcgc   4080 

agttaaaggt ttatctcaat gatgaactga tgggcgtgct gccagtgacc aaagaacagt   4140 

tgggtaaaaa aacgctggcg caaatgccca ttaacccact gtttattagc gacttcaacc   4200 

gtgtacggct ggagtttgtc ggccattatc aggacgtgtg cgaaaaaccg gccagcacca   4260 

cgctttggct ggatgttggg cggagcagtg gactggatct gacctatcag accctgaatg   4320 

tgaagaatga cctgtcacac ttcccggtgc cattctttga cccgagcgat aaccgcacca   4380 

acaccttgcc gatggtcttt gcgggtgcgc cggatgttgg gctgcaacaa gcctctgcca   4440 

ttgtcgcctc gtggtttggt tcgcgttctg gctggcgtgg gcagaacttc ccggtactct   4500 

ataaccaact gccggatcgc aatgccattg tctttgcaac caacgacaaa cggccggact   4560 

tcctgcgcga tcatccggcg gtaaaagccc cggtgattga gatgattaac catccgcaga   4620 

atccttacgt caaactgctg gtggtgtttg gtcgtgacga caaagacctg ttgcaggcag   4680 

cgaaaggtat cgctcagggt aacattctgt tccgtggtga aagcgtggta gtgaatgaag   4740 

tgaaaccgct gctaccgcgt aagccgtacg atgcgccgaa ctgggtacgt accgatcgtc   4800 

cggtcacatt tggcgaactg aaaacctatg aagaacagtt acaatccagc ggtcttgagc   4860 

cagcagcgat taacgtttcg ctaaacctgc cgccggatct ctacctgatg cgcagtaccg   4920 

gcattgatat ggatattaat taccgctaca ccatgccgcc ggtgaaagac agttcgcgga   4980 

tggatatcag cctgaataac cagttcctgc aatccttcaa cctgagcagc aaacaggagg   5040 

cgaaccgcct gctgctgcgg attccggtat tacaaggttt gctggatggc aaaacagatg   5100 

tctctattcc ggcgctgaaa ctgggcgcga ccaaccagct gcgcttcgac tttgagtata   5160 

tgaacccgat gccgggcggt tcggtggata actgtattac cttccagccg gtgcagaatc   5220 

atgtggtgat tggtgacgac tccaccatcg acttctcgaa gtattaccac ttcatcccga   5280 

tgccggatct acgcgccttt gctaacgcgg gcttcccatt cagccggatg gcggatctgt   5340 

cgcaaaccat caccgtgatg ccgaaagcgc ctaacgaagc acagatggaa acgttgctga   5400 

atactgttgg ttttatcggc gcacagacgg gcttcccggc gattaatctg acggtgaccg   5460 

atgatggcag caccattcag ggcaaagatg ccgacatcat gatcatcggt ggtatcccgg   5520 

acaaactgaa agacgataag cagatcgacc tattggtgca ggcgaccgaa agctgggtga   5580 

aaacaccgat gcgccagacc ccgttccccg gcattgtgcc ggacgagagc gatcgcgcgg   5640 

cagaaacccg gtcaacgctg acctcttccg gtgcgatggc ggcggtgatt ggcttccagt   5700 

cgccgtataa cgaccagcgc agcgtgattg cgctgttggc agatagccca cgcggttatg   5760 

aaatgcttaa cgatgcggtg aacgatagcg gcaaacgcgc caccatgttc ggttcggtcg   5820 

cggtgatccg cgagtccggt atcaacagcc tacgtgttgg cgacgtttat tacgtaggtc   5880 

atctgccgtg gttcgagcgc gtgtggtatg cgctggcaaa ccatccgatt ctgctggcgg   5940 

tgctggcggc tatcagtgtg atattgctgg catgggtact gtggcgtctg ctgcgaatta   6000 

ttagtcgtcg tcgtcttaac ccggataacg agtaattgaa gatgaatgtg ttgcgtagtg   6060 

gaatcgtgac gatgctgctg ctggctgcct ttagtgttca ggcagcctgt acctggcctg   6120 

cctgggagca gtttaaaaag gattacatca gtcaggaagg gcgcgtcatc gaccccagcg   6180 

acgcgcgcaa aatcaccacc tccgaagggc aaagttacgg catgttctct gccctggcgg   6240 

ctaacgaccg tgcagctttc gataatattc tcgactggac gcagaacaat ctcgctcagg   6300 

gttctttaaa agaacgtttg cccgcctggc tgtggggcaa gaaagagaac agtaagtggg   6360 

aagtgctgga cagcaattcg gcctccgatg gtgatgtctg gatggcctgg tcgttgctgg   6420 

aggcggggcg tttgtggaaa gagcagcgtt ataccgacat cggcagcgcg ttgctaaaac   6480 

gtatcgcgcg ggaggaagtg gtgacggtgc ctgggctggg ttccatgttg ttaccgggca   6540 

aagtgggttt tgctgaggat aacagctggc gttttaaccc cagctacctg ccgccgacgc   6600 

tggcgcagta tttcacccgc tttggcgcgc cgtggaccac gctgcgcgaa accaatcaac   6660 

gtttattgct ggaaaccgcc ccgaaaggtt tttcgccaga ctgggtgcgc tatgagaaag   6720 

acaaaggctg gcagctaaaa gccgaaaaaa cattgatcag cagctacgac gctatccgcg   6780 

tttacatgtg ggtaggcatg atgcctgaca gcgatccgca aaaagcgcgg atgctcaacc   6840 

ggtttaaacc gatggcgaca ttcactgaga aaaacggtta tccgccggaa aaagtggatg   6900 

tggctacggg gaaagcgcag ggtaaaggac cagtcggttt ttctgccgcc atgctgccct   6960 

ttttacaaaa ccgcgatgcg caggccgttc agcgccagcg cgtggccgat aactttcccg   7020 

gcagcgatgc ctattacaac tatgtgctga ccctgtttgg acaaggctgg gatcaacacc   7080 

gtttccgctt ctcgacaaaa ggtgagttat tacctgactg gggccaggaa tgcgcaaatt   7140 

cacactaaac atattccgct ttccctcggt ctggccgtca tgccgatggt cgaggcagca   7200 

ccaaccgctc agcaacagtt gctggagcaa gttcggttag gcgaagcgac ccatcgtgaa   7260 

gatctggtgc aacagtcgtt atatcggctg gaacttattg atccgaataa cccggacgtc   7320 

gttgccgccc gtttccgttc tttgttacgt cagggcgata ttgatggcgc gcaaaaacag   7380 

ctcgatcggc tgtcgcagtt agcgccgagt tcaaatgcgt ataaatcgtc gcggactacg   7440 

atgctacttt ccacgccgga tggtcgtcag gcactgcaac aggcacgatt gcaggcgacg   7500 

accggtcatg cagaagaagc tgtggcgagt tacaacaaac tgttcaacgg tgcgccgccg   7560 

gaaggtgaca ttgctgtcga gtactggagt acggtggcga aaattccggc tcgccgtggc   7620 

gaagcgatta atcagttaaa acgcatcaat gcggatgcac cgggcaatac gggcctgcaa   7680 

aacaatctgg cgctattgct gtttagtagc gatcgccgtg acgaaggttt tgccgtcctg   7740 

gaacagatgg caaaatcgaa cgccgggcgc gaaggggcct ctaaaatctg gtacgggcag   7800 

attaaagaca tgcccgtcag tgatgccagt gtgtcggcgc tgaaaaaata tctctcgatc   7860 

tttagtgatg gcgatagcgt ggcggctgcg caatcgcaac tggcagaaca gcaaaaacag   7920 

ctggccgatc ctgctttccg cgctcgtgcg caaggtttag cggcggtgga ctctggtatg   7980 

gcgggtaaag ccattcccga actacaacag gcggtgcggg cgaacccgaa agacagtgaa   8040 

gctctggggg cgctgggcca ggcgtattct cagaaaggcg atcgcgccaa tgcagtggcg   8100 

aatctggaaa aagccctcgc actggacccg cacagcagca acaacgacaa atggaacagt   8160 

ctgctgaaag taaaccgcta ctggctggcg atccagcagg gcgatgctgc gctgaaagcc   8220 

aataatcctg accgggcaga acgcctgttc cagcaggcgc gtaatgtcga taacaccgac   8280 

agttatgcag tgctggggct gggcgatgtg gcgatggcgc gaaaagatta tcccgccgcc   8340 

gaacgttatt atcagcagac cttgcgtatg gacagcggca acactaacgc cgtgcgcggg   8400 

ctggcaaata tttaccgcca gcaatcgcca gaaaaagctg aagcgtttat cgcctcgctc   8460 

tctgccagtc agcggcgtag cattgatgat atcgaacgca gcctgcaaaa cgaccgtctg   8520 

gcacagcagg cagaggcact ggaaaaccag ggcaaatggg cgcaggcggc agcacttcag   8580 

cggcaacgac tggcgctgga ccccggcagc gtatggatta cttaccgact ttcgcaggat   8640 

ctctggcagg ccggacaacg cagccaggcc gatacgttaa tgcgcaatct ggcgcagcag   8700 

aagtcgaacg acccggagca ggtttacgct tacgggctgt acctctctgg tcatgaccag   8760 

gacagagcgg cgctggcgca tatcaatagc ctgccgcgtg cgcagtggaa cagcaatatt   8820 

caggagctgg ttaatcgact gcaaagcgat caggtgctgg aaaccgctaa ccgcctgcga   8880 

gaaagcggca aagaggcaga agcggaagcg atgctgcgcc agcaaccacc ttccacgcgt   8940 

attgacctca cgctggctga ctgggcgcaa caacgacgtg attacaccgc cgcccgcgct   9000 

gcatatcaga atgtcctgac gcgggagcca gctaacgccg acgccattct tggtctgacg   9060 

gaagtggata ttgctgccgg tgacaaagcg gcggcacgta gccagctggc gaaactgccc   9120 

gctaccgata acgcctcgct gaacacacag cggcgcgtgg cgctggcaca ggcgcagctt   9180 

ggcgataccg cagcagcgca gcggacgttt aataagttga tcccgcaggc aaaatctcag   9240 

ccaccgtcga tggaaagcgc gatggtgctg cgtgatggtg cgaagtttga agcgcaggcg   9300 

ggcgatccaa cgcaggcgct ggaaacctac aaagacgcca tggtcgcatc cggtgtgact   9360 

acgacgcgtc cgcaggataa cgacaccttt acccgactga cccgtaacga cgagaaagat   9420 

gactggctga aacgtggcgt gcgcagcgat gcggcggacc tctatcgcca gcaggatctt   9480 

aacgtcaccc ttgagcacga ttactggggt tcgagcggca ccggtggtta ctccgatctg   9540 

aaagcgcaca ctaccatgtt gcaggtggat gcgccgtatt ctgacgggcg gatgttcttt   9600 

cgcagtgatt tcgtcaatat gaacgtcggc agtttctcca ctaatgccga tggcaaatgg   9660 

gatgacaact ggggcacctg tacattacag gactgtagcg gcaaccgcag ccagtcggat   9720 

tccggtgcca gcgtggcggt cggctggcga aatgacgtct ggagctggga tatcggtacc   9780 

acgccgatgg gcttcaacgt ggtggatgtg gtcggcggca tcagttacag cgatgatatc   9840 

gggccgctgg gttacaccgt taacgcccac cgtcggccca tctccagttc tttgctggcc   9900 

tttggtgggc aaaaagactc cccgagcaat accgggaaaa aatggggtgg cgtacgtgcc   9960 

gacggtgtgg ggctaagtct gagctacgat aaaggtgaag caaacggcgt ctgggcatcg  10020 

cttagtggcg accagttaac cggtaaaaat gtcgaagata actggcgcgt gcgctggatg  10080 

acgggctatt actataaggt cattaaccag aacaatcgcc gcgtcacaat cggcctgaac  10140 

aacatgatct ggcattacga caaagatctg agtggctact cactcggtca gggcggttac  10200 

tacagtccgc aggaatacct gtcgtttgcc ataccggtga tgtggcggga gcgcacggaa  10260 

aactggtcgt gggagctggg tgcgtctggc tcgtggtcgc attcacgcac caaaaccatg  10320 

ccgcgttatc cgctgatgaa tctgatcccg accgactggc aggaagaagc tgcgcggcaa  10380 

tccaacgatg gcggcagcag tcagggcttc ggctacacgg cgcgggcatt acttgaacga  10440 

cgtgttactt ccaactggtt tgttggcacg gcaattgata tccagcaggc gaaagattac  10500 

gcacccagcc atttcctgct ctacgtacgt tattccgccg ccggatggca gggtgacatg  10560 

gatttaccgc cgcagccgct gataccttac gccgactggt aagttttcag atagcgcctc  10620 

tcttaatgcc gctgcgatcg ggtatactcg ggcggcaatc tgggatttcc ggggggagac  10680 

aatttgcgcg taagtcgctc gttaacaatc aagcagatgg caatggtggc agccgttgtc  10740 

ctggtgttcg tttttatttt ttgcaccgtt ttgctgttcc atctggtcca gcagaatcgc  10800 

tataacacgg ctacgcaact ggaaagcatt gctcgctctg tccgcgaacc cttatcttca  10860 

gctattttga aaggcgatat tcccgaagcg gaagctattc ttgccagcat taaaccggca  10920 

ggcgtggtca gccgtgccga tgtagtgctg cctaaccagt tccaggcgct gcgtaaaagt  10980 

tttattccag agcgcccggt gccggtaatg gttactcgcc tgtttgagct accggttcaa  11040 

atctcgctgg gcgtttactc gctcgaacgt ccggcaaacc cgcagccaat tgcctatctg  11100 

gtactacagg cggattcctt ccgtatgtat aagttcgtga tgagcaccct ctcaacgtta  11160 

gtgaccattt acttactttt gtcgcttatc ctgaccgtcg ccatcagctg gtgcattaac  11220 

cgcctgattt tgcatccgtt acgcaatatt gctcgcgaac ttaacgccat cccagccaag  11280 

gagcttgttg gtcaccaact ggcattaccg cgtctgcatc aggacgatga aatcggtatg  11340 

ttggtgcgca gttacaacct caaccagcaa ttgctgcagc gccattatga agaacagaac  11400 

gaaaatgcga tgcgcttccc ggtgtcggat ttgccgaaca aagccttgct gatggagatg  11460 

ctggagcagg ttgtcgcgcg taaacaaacc accgcgctga tgatcatcac ctgtgaaacc  11520 

ctgcgtgata ctgcgggcgt gctgaaagag gcgcaacgag aaattctgct gctgacgctg  11580 

gtggaaaaac tcaaatcggt actgtcgcca cgtatgatcc tcgcgcagat tagcggttat  11640 

gactttgctg tcattgccaa cggtgtacag gaaccgtggc acgcaatcac cttaggtcag  11700 

caagtgctca ctatcatgag cgagcgcctg ccgattgaac gtattcaact ccgtccgcac  11760 

tgtagcattg gcgtggcgat gttctacggc gatctcaccg ccgaacagct ttacagtcgc  11820 

gctatttctg cggcatttac cgctcgccat aaaggcaaga atcagattca gttctttgat  11880 

ccgcagcaga tggaagccgc ccagaagcgg ttgacggaag agagcgatat ccttaatgca  11940 

ctggaaaatc atcagtttgc tatttggtta cagccacagg tcgagatgac cagcggtaaa  12000 

ctggtcagtg cggaagtgtt actgcgtatc cagcaaccgg atggcagttg ggacctgccg  12060 

gatggcttaa tcgatcgcat tgagtgctgt gggctgatgg ttaccgtcgg tcactgggtg  12120 

ctggaagagt cctgtcgatt gcttgcagcc tggcaagagc gcggcattat gctgcccttg  12180 

tcggtaaacc tctctgcgct gcaactgatg cacccgaata tggtggcgga tatgctggaa  12240 

ctgttaaccc gctatcgcat tcagccggga acactgattc tggaagtgac agaaagccga  12300 

cgtattgacg accctcatgc tgcggtggca atcctccgtc cgctgcgcaa tgccggagtt  12360 

cgggtggcgc tggatgattt cggcatgggc tacgcagggc tgcgtcagct gcagcatatg  12420 

aaatcgttgc caatcgacgt actgaaaatc gacaaaatgt ttgttgaagg cttgccggga  12480 

gatagcagca tgattgctgc aattatcatg ctggcgcaga gcctgaactt acaaatgatt  12540 

gccgaaggcg tggagactga agcacaacgc gactggctgg caaaagcggg cgttggtatt  12600 

gcccagggct tcctttttgc tcgcccactc cctattgaaa tcttcgaaga gagttacctg  12660 

gaagaaaagt agctacccca aactgattac aaaactttaa aaagtgctgg tttgtgcgag  12720 

ccagctcaaa ctttttaacc tttttgtttc aattatgatc caggtacatt tctgtgatgt  12780 

tgtctgggtg ttattttaag gccgcaggta ccccataacc ttacaagacc tgtggtttta  12840 

ctaaaggaca ccctatgaaa acctctctgt ttaaaagcct ttactttcag gtcctgacag  12900 

cgatagccat tggtattctc cttggccatt tctatcctga aataggcgag caaatgaaac  12960 

cgcttggcga cggcttcgtt aagctcatta agatgatcat cgctcctgtc atcttttgta  13020 

ccgtcgtaac gggcattgcg ggcatggaaa gcatgaaggc ggtcggtcgt accggcgcag  13080 

tcgcactgct ttactttgaa attgtcagta ccatcgcgct gattattggt cttatcatcg  13140 

ttaacgtcgt gcagcctggt gccggaatga acgtcgatcc ggcaacgctt gatgcgaaag  13200 

cggtagcggt ttacgccgat caggcgaaag accagggcat tgtcgccttc attatggatg  13260 

tcatcccggc gagcgtcatt ggcgcatttg ccagcggtaa cattctgcag gtgctgctgt  13320 

ttgccgtact gtttggtttt gcgctccacc gtctgggcag caaaggccaa ctgattttta  13380 

acgtcatcga aagtttctcg caggtcatct tcggcatcat caatatgatc atgcgtctgg  13440 

cacctattgg tgcgttcggg gcaatggcgt ttaccatcgg taaatacggc gtcggcacac  13500 

tggtgcaact ggggcagctg attatctgtt tctacattac ctgtatcctg tttgtggtgc  13560 

tggtattggg ttcaatcgct aaagcgactg gtttcagtat cttcaaattt atccgctaca  13620 

tccgtgaaga actgctgatt gtactgggga cttcatcttc cgagtcggcg ctgccgcgta  13680 

tgctcgacaa gatggagaaa ctcggctgcc gtaaatcggt ggtggggctg gtcatcccga  13740 

caggctactc gtttaacctt gatggcacat cgatatacct gacaatggcg gcggtgttta  13800 

tcgcccaggc cactaacagt cagatggata tcgtccacca aatcacgctg ttaatcgtgt  13860 

tgctgctttc ttctaaaggg gcggcagggg taacgggtag tggctttatc gtgctggcgg  13920 

cgacgctctc tgcggtgggc catttgccgg tagcgggtct ggcgctgatc ctcggtatcg  13980 

accgctttat gtcagaagct                                              14000 

 
           
             27  
             13288  
             DNA  
             Pseudomonas fluorescens  
           
            27 

atatgacgag tctagagggt aagttcaatt atcattttag cggttggtta acatttatcg     60 

aaaagtgatc taatgatcat agtatggccg ttagtaaatt catgatggaa agtatacgtc    120 

agtaatcgcc cgtgtatcac gatagccaag catctaagcg cgcgtattgt gtgtttgcct    180 

gacgaaatac tggtacatct cacggcttgc gtgtcttgac tcttgaagaa ggatcttcat    240 

tcagatttcg cacttcaggt tggcgttttt gtcaggccgc tttgatcgat gcaacttttc    300 

cgggctgtct gcgtcgacat ggaccactga ttggaaaccg ggtgtcttac gtgcattctc    360 

cagcatcagg ctgagccgat catccggaat catggcgcca gcccatatgt aattgatttc    420 

cttcttgatc ggcgtaacgg ccgtgggaac gggggcaagg gctgcgtgaa ctgtttcaaa    480 

ggtggttggc ttgggcagtg tcgccttgac ccgcaagaac tcgtccaggc tgccactttt    540 

ccactcgtag gggcctgttc gccaaaaagg ggcgcccagg gttttaccgt cgcctagctt    600 

gcgccaaacg tcattggcta catcaaactc tactaaaacg caaaaattgg cataaagcct    660 

tgtcttgcca tcatcactaa gtacgcggtt gacttcgtcc attgggactt catagctttt    720 

ataaggcatt ccctcgacac cgcccggcag ccccaggcgt tgccattgat cggcgccggt    780 

tttgtttaat atgaattcag tttgtcggta ggtcttgggg ttgatgacac gaacctgcac    840 

cgaactcatg gggctaccgc ccgaagcttc tcgaacctga tagacagaag ccttgccaga    900 

cttgtcgacg tggcggatgt attgactctg gccatcgagg ctgtggtaaa ccccttggct    960 

atccgggctc aggccgacga ggcgcgaggg tgaaactgtg tatttttcga ccactgcgag   1020 

ttcacccggc aggccgggtg gatgcttcaa cttgaacgcg cgagcagccg ttgaaagcat   1080 

ttggcgcgtg ccttgaccca aaccctcggc cagcgcgcca aggccgtccg ctgggttaag   1140 

agcgccgact gccgcggcgc cgattatttt ggccccggtg aacagttttg ttccgagttt   1200 

tccagtgagc ttggtcgcct ggctgaccgc cttgcccacc ggaataacaa aacccagcag   1260 

gtccagagcg aagtccccaa cggcccccca tacgtcgcct tttactgcct tttcaatacc   1320 

gcttttcagg ggcaggaagt ccatgacggc gcccagttca tccaccaact gtttttcttg   1380 

ctcttcaaca gcattggttc cactggcgta ggtcttgagc gcgtcgcggc catagaggac   1440 

aaacttttcg atctcgttgg ccaggtgttg ctcacgctca ctggcccatg gcgctttagg   1500 

ggaacggtct tgcccggtct tttctcccca tgcctttgga aaagaaaaag gcaccggatc   1560 

aatggaaagg cctgaagaca cgacctcctt tggcggcgcg ccggttttgt aggcgtccaa   1620 

gtcaatgtcg agcgacttgc ctttggcctt ttcaagaaag tagtcatccg ggtcatcgta   1680 

gccacgtagg ggcgtttgat gaacgctgct gccccatagc tgatggttgc ggcgaacgag   1740 

cccctgcatc ggcaacactt cgtaaatccg cacgtcacct ttgtttttta cacacatcag   1800 

tagaccgtga cgtgctctgg acgcgtcgac ttgagcgggt gtctcagtgt cgccgttggc   1860 

ttcgcgcaca gagtggaagg tgatgtcgcc gaccgcgata tgttggcgtt catcaagagg   1920 

cagttgggaa aactggtatt gcatcgcggt gtttatgcct gtcctcaatc ccttgatgta   1980 

ggcatcgaac tgctcattga aaatggccgt gatgtccgga aggggtctga cccccgtgcg   2040 

gcgggtcggg ctcagtacga cgctctggtc aacgatacgg tccatcttcc ctgccgtata   2100 

aatgtctacc agcgcatgct tggtctcact gccgtcaagg ttgacgatgg gggtcagcgg   2160 

ctgctcgaag tcatagtcgc catatacctt cttcaattct gcgcaggcca atgccctacg   2220 

cgtaggcaga aaggcggtga ggcctttatt ggcagtgcct aatgtttcta gcaatgcgtt   2280 

gaatttgtct cgcgcaacgg tgatctgttc gggcgtaaac gcatcatcgt tagccttatt   2340 

caggacgtca ttggcgatgg cccaatcgat gacagcatgt gtctgagccc ccgcctcaat   2400 

gccagcatcc ttaagcgtga ccgggcttct gttgccaaaa ctcatgactt ggccaaacgt   2460 

catgttacgg gtggcgcccg gctccagcgc ctcaatgcgc gcgacagcgg tgctcaggct   2520 

ggcccaggtg tgggacccat acaccaagcc gccagggagg tccttgacca ggaacgcagg   2580 

tgctgcactg gcgagcaata gctgtgccgc cgcaggggcc aggtcgtgat cgactcggcc   2640 

ttgctgctta agatgctcga cgagtcgcgc tgttataacg gcaggtgaca agccccggtg   2700 

agctgcctgg gccaggtcgt aaccggcgac gacgttgcgt tgggtgccca cggcgggatc   2760 

caggtcgagc gccagcgcgg tcatcgccca gtcgctggct gtgctggttg accaggtgtc   2820 

cgcgaacgcg cttcgcaggc gcttacccaa cgcctgtgcc tgcggactag caatcagagc   2880 

ctgcaggcgt tgcgcggggc tatcagcgtt ggtgagatcg ggagtttttt tagccaggtg   2940 

gttgagaagg caggtgcccc cctccaaccc atttcgtgcc tcttcactga cgatgttctg   3000 

gacgcgttcg cgctcgtcac gtccgagttg gcgcgtgtcc gacagcagtc cccagtgcct   3060 

cgagttgtca ggcggttgtg gcaacgggtc tgacagaacg gcaataaggt tcgacagctc   3120 

ggccttgtcc tggggaatca tcaagccgtg cgcatgcaaa aacggctcaa gcttcaaacc   3180 

gctttccggc atgcctgggt aagtcttcgc gaatacggaa tccgggtgga cggggacacg   3240 

gtgcgagtcg atgtgttcgg ccagcgtcgt gtttgaattc gttgaggcca gcacactcat   3300 

acgggtgagc gtatggataa ggcttaggcg gttgcgtctg tcacccaggt tttgtttctg   3360 

aagctcgagg gcttcagggg tgcgtgtatg gggaggtctt aacccatccg atgggtcgag   3420 

tggtcggaaa ccctggcctt cattcaattg tgcaacacgc gttgccacct gggcccggct   3480 

gaggttgtcg aggttttccc catagaaccg cccgataagc tcaaagggcg cgctgtgtac   3540 

aggtagcatt gcgggattcc tgatactgaa ggtatcgggc gcgatcacct tgccggcgtc   3600 

gagtatgggg ccggccaccg ccgcccagcg cgcattgttg ttgagggtaa aactgaccac   3660 

gctctggtcg gcttttcgcc tggcgtgtaa ggcgcctttg gagggaatga tctcgacagt   3720 

gctgctatcg agaccctccg aaagcatcca ggcggtgaag tccgggctat tcaacgtctc   3780 

ccgaaagtat tgccaccatt ttccgaacgt cgagtcgggg ggaatgccgg caacaatcac   3840 

ctctcgttgg ctgtccccgc cgcgcgcgcg tgccagcgcg tcgccgtagt aacctgccag   3900 

gagcttgtcc gggtaaccca tcgctaaagc attgcctgcg gcgatgctgg ctttcgctct   3960 

atttgaaggc agcagactca acggcaacgg cgactttgct ggcgtcttgg cggtctcggg   4020 

cacggaatct gtggaggctc tgtcgctgcg atgctggctg aatgggttgc cggtggtagg   4080 

gggggcgatc cgtgtgtttg gcgggggcgt aatggctagc ggggtagagg gggcggaaat   4140 

attcaatggt tatgctcctt gcagggatag gcgctgaagc gccaggcgtg ggcaggcagt   4200 

tctacgccct cgtcgctgtc tcgcagagtg cgtttgtttt ttcgggtgtt tccgagcggc   4260 

tccagcctgc gtaaaacctg ggtgcgtggg ttgtatcttc tattgtccac tgtaaaaaaa   4320 

agaacacggc ttgacgttgc ggaggtgggg ccccgccttc cgatttcgga ttcgacccgt   4380 

cctgatattc agcgaggagc acgtcagtgg aggcggtgtt gagggaggag ggggcggcaa   4440 

tcaccccgcg aggatttttt gcctgtggag cgttctatcc gataacccat tgataaacga   4500 

tggcactgcc tcagctatac tcccctccat ttgaatggcc cttcgaggaa ttactgtgaa   4560 

gaactggacc ttgcgccaac gcatcttggc aagttttgcg gtcattatcg ccatcatgct   4620 

gttaatggtc gtggtctcct attcacggtt gctgaagatc gaaaccagcc aggaagccgt   4680 

ccgggacgac gcggtgccgg gcgtttacct cagctcgatg atccgcagcg cctgggtcga   4740 

cagctatttg cagaccatcg acatcatcgg cctgcgcgac gacaaaacct tcaccaatac   4800 

cgataagaac gactacaagt cgttcgaagc gcgtattgaa cagcagatgg ccaactacga   4860 

aaaaaccatt catggccaag ctgaccgcat ggagttcgat aacttcaagg cggcgcacat   4920 

caactacaac aaagtgctgg cccaggtgct ggaacgcgtt gaagccaatg acctgccggg   4980 

cgcgaatcaa ttgctcgagg agcaattgac gccgatctgg accgaagggc gcatgaagct   5040 

caacgacatc attactgaaa acaagaacgt gtctgaccgg gcgacggcgg cgatcgacga   5100 

ggcggtactg tcggccaaga tcagcatggc cgtgtcgctg ctcatcgcca tcttggctgc   5160 

cgggctgtgc ggcctgttgc tgatgcgcgc gatcatggcg ccgatgcaac gcatcgtcga   5220 

tatcctcgaa accatgcgcg acggcgacct gagcaaacgc cttaacctgg agcgcaagga   5280 

cgagttcggc gccgtcgaaa ccggctttaa cgacatgatg accgagctca cggccctggt   5340 

gtcccaggcc cagcgctcgt cggtgcaggt caccacctcg gtgaccgaaa ttgccgccac   5400 

gtccaagcag caacaggcca ccgcgactga aacggccgcg acgaccactg agatcggtgc   5460 

tacgtcgcgc gaaatcgcgg ccacctccaa ggacttggtt cgcaccatga ccgaagtgtc   5520 

caccgccgcc gaccaggcgt cggtggctgc cggctccggc cagcaaggac tggcgcgcat   5580 

ggaagagacc atgcactcgg tgatgggcgc ggccgacctg gtcaacgcca agctggcgat   5640 

cctcaatgag aaggccggca acatcaacca ggtggtggtg accatcgtca aggtggccga   5700 

ccagaccaac ctgctgtcgc tcaacgccgc catcgaagcg gaaaaagccg gcgagtacgg   5760 

tcgcggtttt gccgtggtcg ccaccgaagt acgccgcctc gcggaccaga ccgccgtggc   5820 

tacctatgac atcgagcaga tggtgcgcga gatccagtcc gcagtgtctg cgggggtgat   5880 

gggcatggac aagttctccg aagaagtgcg ccgcggcatg ttcgaagtgc agcaagtggg   5940 

cgagcagttg tcgcagatca tccaccaggt acaggcgctg gcgccgcggg tgttgatggt   6000 

caacgaaggc atgcaggccc aggccaccgg cgccgagcag atcaaccacg ccctcgtgca   6060 

actgggcgat gccagcagcc agacggtgga gtccctgcgc caggccagct ttgccattga   6120 

tgaactgagc caggttgcgg tgggtctgcg cagcggcgtg tcgcgtttca aagtctgatg   6180 

agcgaactcg cggctaaacg cggcgccgtc ccggcagcga aaaaggcgtt gttcctggtg   6240 

ttccatatcg gtcaggaacg ctatgccctc aaggctaccg aagtggccga agtgctgccg   6300 

cgcctgcctt tgaaacccat tgcccatgca ccgctgtggg tcgccgggat ctttgcccat   6360 

cgtggggcgc tggtgccggt cattgacctc agcgccttga ccttcggcaa cccggcccag   6420 

gcccgcacca gcacgcggct ggtgctggtc aattaccaac ccgacgccgg gtcccaagcg   6480 

cgttggctgg gactgatcct ggagcaggcc accgacaccc tgcgttgcga ccccgccgag   6540 

ttccagccct acggtctggc caaccgccag gcaccctacc tggggccggt gcgcgaagat   6600 

gcgctgggcc tgatgcaatg gatcggtgta aacgacctgc tgaccgatga cgtgcgcgcc   6660 

gtgctgtttt ctgccgagct gagcgtatga gcaacgaccc gcgttttttt gcctttctta   6720 

aagagcgcat cggcctggac gtcgcgtcgg tgggcgaagc catcatcgag cgcgccgtgc   6780 

gccagcgcag ccagatcgtc caggcaccca cgccgggcga atactggcag cacctgcaaa   6840 

gctcccagga cgaacagcaa gcgctgatcg aagcagtgat cgtccccgaa acctggtttt   6900 

tccgttaccc cgaatccttt gcgaccctgg cgcgcctggc gaaggcccgc ctggtcgata   6960 

tcaagcagat gcgcgcattg cgtatcctca gcctgccgtg ctccaccggc gaagaaccct   7020 

attcgattgc catggcgctg ctcgatgctg gcctcgcgcc gcatcagttc aaggtacagg   7080 

ggatggacgt cagcccgctg tcggtggagc gtgcgcgacg cggggtgtac ggcaagaact   7140 

cctttcgtgg cggcgatatt gccttccgtg accgccactt cactgaatac ggcgacggct   7200 

tccacatcgc cgatcgggtg cgcgaacaag tgcgcctgca agtcggcaac ctgcttgacc   7260 

cggcgctgct ggttaatgag gccgcctatg acttcgtgtt ctgccgcaac ctgctgatct   7320 

acttcgacca gcccacccag aaacaagtct tcgacgtgct caaaggcctc acccacgtgg   7380 

acggtgtgct gtttatcggc ccggccgaag gcagcctgct ggggcgccac ggcatgcgtt   7440 

cgattggggt gccgcagtcc tttgcgttca gccggcaggg ctcgccgcag ctgcccgagc   7500 

cagcgtttat accgacgccg gctcccacgc ctccgcgcag cacggcgccg atttcagcta   7560 

aaccacggcc gttcagcacc gtcagcgccc acgttttacc gatcaaggct acgccctccg   7620 

atgcaggcac cttgctcagc cggatcgcca ccctggccaa cgaaggcaaa agtgccgagg   7680 

cccgggccgc gtgtgaagac tatttgaaca gccacccgcc ggccgcccag gtgttttact   7740 

ggctggggct gctcagcgac gtggccggca gtgccctgga agcccagggc tattatcgaa   7800 

aagccttgta cctggaaccc cagcacccgc aggccttgat gcacttggcc gcgttgctcg   7860 

agtcccgggg cgacagtgcg ggggcgcgcc gtctgcaggc gcgagccgcc cgtagcgagc   7920 

gagccgacag tgagtccaaa ccatgagtaa caccgaggcg ctcgacacca ccggcctgga   7980 

cctgaccctg gccgacactc aagccatcga cgactgctgg aaccgcatcg gtatccatgg   8040 

cgataagtcc tgcccactac tggctgacca tatccattgc cgcaactgct cggtgtattc   8100 

cgccgccgcc acgcgtctgc tggaccgcta cgccttgcag caggacgacc gacgcccgca   8160 

ggcggccgaa gtcgacacgg aggtggtcac ccgttcgctc ttgatgttcc gcctcggcga   8220 

agaatggctg ggcatcgcta cacgctgcct ggtggaagtc gcgccgttgc aaccgatcca   8280 

ttccctgccg catcaacgtt cccgcgcctt gctcggcgtg gccaacgtgc gcggcgcgct   8340 

ggtggcgtgc ttgtcgctgg tggaactgct gggcctggac gccaccagca gcggcgccac   8400 

cggcgggcgc atcatgccgc gcatgttgat cattgccgcc caggatggcc cggtggtggt   8460 

gccggtggat gaagtcgacg gcatccatgc catcgatgag cgcaccttga aggccgcatc   8520 

ggcgtccggc acccaggcca gcgcgcgctt tacccagggt gtattgccgt ggaaaggccg   8580 

cagcctgcgc tggctggacg aggcgcaatt gttgtccgcc gtgacccgga gcctctcatg   8640 

acccccgacc agatgcgtga tgcctcgctg ctggaattgt tcagcctgga agccgatgcg   8700 

cagacccagg tgttgagcgc gggcctgctc gccctggaac gcaacccgac ccaggcggac   8760 

cagctcgaag cctgcatgcg cgcggcgcac tcgctcaagg gcgccgcgcg catcgttggg   8820 

gtggatgccg gggtcagcgt gtcccacgtc atggaggatt gcctggtcag cgcccaggaa   8880 

aaccgcctgt acctgcaacc cgaacatatc gatgcgctgc tgcagggcac tgatctgctg   8940 

atgcgcatcg ccacgccagg caatgacgtc gggccggcgg atgtcgaggc ctacgtcgcg   9000 

ttgatggagc gtctgctgga tccgtcccag gcgcctgtca acgtcgcgcc gtcgccggag   9060 

ccagcgcccg tcgtcgaaga actgccgccg gaacccgaac ccgcgccgcc ggtgaacagc   9120 

gagccgccgc gtcaaggcaa gcgcatgacc gaaggcggcg aacgcgtact gcgcgtcacc   9180 

gccgagcgct tgaacagcct gctggacctg tcgagcaaat ccctggtgga aacccagcgg   9240 

ctcaagccgt acctggccag cctgcaacgc ctcaagcgca ttcaaagcca ggggatacgg   9300 

gcgctggata cgctggacgg gcaactcaag acccaggtcc tgagcctcga ggcccaggaa   9360 

gccctggccg acacgcgccg tttgttgagc gaagcccagg ccttgctggc ggaaaagcac   9420 

gccgagctgg acgagttcgg ctggcaggcc gggcagcgcg cccaagtgtt gtatgacact   9480 

gcactggcct gccgcatgcg cccgtttgcc gatgtattgg ctggacaagt gcgcatggtg   9540 

cgcgacctcg gccgcagcct cggcaagcaa gtgcgcctgg agatcgaggg cgaaaagacc   9600 

caggtcgacc gtgacgtact ggaaaaactt gaggcgccgc tcacccattt gttacgcaat   9660 

gccgtcgacc acggcatcga aatgcctgag caacgcctgc tggcgggcaa gccggctgaa   9720 

ggcttgatcc gcctgcgggc ctcccatcag gccggtttgc tggtgctgga actcagtgat   9780 

gacggcaatg gcgtcgacct ggagcgcctg cgcggcacta tcgtcgatcg gcacctgtcg   9840 

ccggtcgaaa ccgccctgcg cctgagcgaa gaagagttgc tgacgttcct gttcctgccg   9900 

gggttcagcc tgcgtgacac ggtcaccgaa gtgtccgggc gcggcgtggg cctggacgcg   9960 

gtgcagcaca tggtccgcca actgcgcggc gcggtggtgc tggagcagac ggcggggcag  10020 

ggcagtcgtt tccaccttga ggtgccgttg accctgtcgg tggtacgcag cctggtggtg  10080 

gaagtcggtg aggaagccta tgcgttcccg ctggcgcata tcgaacgcat gtgtgacctc  10140 

gcgcccgatg acatcgtgca actggaaggt cgccagcatt tctggcacga gggccggcat  10200 

gttggcctgg tcgccgccag ccagttgttg cagcgcccgg cggggcagag tccgtcagaa  10260 

acgctgaaag tggtggtgat ccgcgagcgc gatacggtgt acgggattgc cgtggagcgc  10320 

tttatcggtg agcgtacgct ggtggtgttg ccgctcgatg atcgcctggg caaggtccag  10380 

gatatttccg ccggtgcctt gctggatgat ggctcggtgg tgttgatcgt cgacgttgaa  10440 

gacatgttgc gttcggtgga caaactgctg aacaccggcc gattggaacg tattgcgcgg  10500 

cgcagccaac aaaccaccga ggcaccgcgt aagcgcgtgc tggtggtcga tgactcgctg  10560 

accgtgcgtg agctgcaacg caaattgctc cttaatcgtg gttatgaagt ggccgtggcg  10620 

gtcgatggca tggacggctg gaacgccttg cgctccgaag actttgacct gttgatcact  10680 

gatattgata tgccccgcat ggacggtatt gaattggtca cactcttgcg ccgtgacagt  10740 

cgcctgcaat cgttgccggt gatggtggtg tcctacaaag atcgcgaaga agaccgacgt  10800 

cgaggactcg acgccggtgc ggactattat ttagccaaag ccagtttcca cgatgacgcc  10860 

ctgctggacg ccgtggtgga actgatcgga ggcgcacggg catgaggatt gcgatcgtca  10920 

atgacatgcc cctggcagtg gaggccttgc gccgcgcctt gagcttcgag cctgcgcacc  10980 

aagtggtgtg ggtggccagc aatgggctgg aagcggtgca acgctgcgcc gaactgacgc  11040 

cggacctgat cctgatggac ctgatcatgc cggtgatgga cggcgtggaa gccactcgcc  11100 

agatcatggc cgagacgccg tgcgccatcg ttatcgtgac cgtcgaccgc caggccaacg  11160 

tgagccgggt gttcgaggcc atgggccacg gcgccctgga cgtggtggac accccgccgc  11220 

tcggcgtggg caaccccaag gatgcggcgg cgccgttgct gcgcaagatc ctcaatatcg  11280 

gctggctgat cggccagcgc ggcacccgcg tgcgcgccga aaccctgccg gcgcgcgcat  11340 

ccggcaaacg tcaaagcctg gtggctatcg gctcctcggc gggtggtccg gctgccctgg  11400 

aaatcctgct caagggttta cctcgcgact ttccagccgc catcgtgctg gtgcagcatg  11460 

tggaccaagt gttcgcggcg ggcatggccg agtggctgag cagcgcctcg ggcctgccgg  11520 

tacgcctggc ccgcgaaggc gagccgccgc aaagcggcgt ggtgctgctg gccggcacca  11580 

accaccacat tcgtttattg aagaatggca cgctagccta tacggcagag ccggtgaacg  11640 

aaatttaccg gccatcgatc gatgtgtttt tcgaaagcgt ggccagccac tggaatggcg  11700 

atgccgtcgg tgtgctgctg accggcatgg ggcgcgacgg ggcccagggc ctcaaattgc  11760 

tacgtgaaca aggttatttg accatcgccc aggatcagca aagctcggcg gtgtatggca  11820 

tgcccaaagc ggcggcggcg atcgatgctg ctgttgaaat tcgcccactg gatagaattg  11880 

cgccccggtt gctggaggtc tttgccaaat gaacatgacc tctcgcggct ggcttgcagg  11940 

cagtaattca ggtgactgca catgaatgat ttacagatcg acgacatcaa gaccgacgaa  12000 

aacgccgcca tggtgttgct ggtcgacgac caggccatga tcggtgaagc cgtgcggcgt  12060 

ggcctggccc atgaagaaaa tatcgacttc cacttctgcg ccgacccaca ccaggcgatt  12120 

gcccaggcga tccgtatcaa gccgaccgtt atcctgcagg atctggtgat gccaggtctg  12180 

gacggcctga cactggtgcg cgagtaccgc aaccacccgg ccacgcagaa catcccgatc  12240 

atcgtgcttt ccaccaagga agacccgctg atcaagagcg cggcgttttc ggccggggcc  12300 

aacgattatt tggtcaagct gccggacaac atcgagctgg tggcgcgcat ccgctatcac  12360 

tcgcgctcct acatgaccct gttgcaacgg gatgcggctt atcgcgcgtt gcgggtcagc  12420 

cagcagcagc tgttggacac caacctggtg ctgcaacggc tgatgaactc cgatggcctc  12480 

acggggctgt ccaaccgtcg ccatttcgac gagtacctgg aactggaatg gcgccgtgcc  12540 

atgcgtgatc agactcagct gtcgttgttg atgattgatg tggatttctt caagacctac  12600 

aacgatagct ttgggcatgt cgaaggtgac gaggctttgc gcaaggttgc ggcgaccatt  12660 

cgtgaggcca gcagtcggcc ttcggatttg ccggcgcgct atggggggga ggagtttgcc  12720 

ctggtgctgc ctaatacctc gccgggcggg gcgcggttgg tggctgagaa gctgcggatg  12780 

gcggttgccg cgctgaaaat tccgcacatt gcgccgactg aggggtcgag tctgaccatc  12840 

agtattgggc tgtcgaccat gacgccgcag caggggacgg attgtcggca ggtgatagtg  12900 

gcggcggata aggggttgta tacggctaag cataatgggc gcaatcaggt ggggattgag  12960 

tagggctgag tgcttgtgta catatccgtt gctgcggtca cggccactaa tggttccgct  13020 

cttacagcag ggtcactttt tgaaaagcgc aaaaagtaac caaaaacgct tcgccccaac  13080 

actcggcacc tcgcctagga ctcggtgtac cctcactcag gattcaatac tgcgttcagc  13140 

caacgtgttt gacggagcgc cttagatcaa aagcaaaaac gcggcggcct taaaacccac  13200 

cgaggttggg gagcattact gggtaaaaat gtgggagggg gcttgacctg gcggtgtgta  13260 

gtgatcgatc tgtggctgta cactgcca                                     13288 

 
           
             28  
             547  
             PRT  
             Pseudomonas fluorescens  
           
            28 

Met Ala Leu Arg Gly Ile Thr Val Lys Asn Trp Thr Leu Arg Gln Arg 
  1               5                  10                  15 

Ile Leu Ala Ser Phe Ala Val Ile Ile Ala Ile Met Leu Leu Met Val 
             20                  25                  30 

Val Val Ser Tyr Ser Arg Leu Leu Lys Ile Glu Thr Ser Gln Glu Ala 
         35                  40                  45 

Val Arg Asp Asp Ala Val Pro Gly Val Tyr Leu Ser Ser Met Ile Arg 
     50                  55                  60 

Ser Ala Trp Val Asp Ser Tyr Leu Gln Thr Ile Asp Ile Ile Gly Leu 
 65                  70                  75                  80 

Arg Asp Asp Lys Thr Phe Thr Asn Thr Asp Lys Asn Asp Tyr Lys Ser 
                 85                  90                  95 

Phe Glu Ala Arg Ile Glu Gln Gln Met Ala Asn Tyr Glu Lys Thr Ile 
            100                 105                 110 

His Gly Gln Ala Asp Arg Met Glu Phe Asp Asn Phe Lys Ala Ala His 
        115                 120                 125 

Ile Asn Tyr Asn Lys Val Leu Ala Gln Val Leu Glu Arg Val Glu Ala 
    130                 135                 140 

Asn Asp Leu Pro Gly Ala Asn Gln Leu Leu Glu Glu Gln Leu Thr Pro 
145                 150                 155                 160 

Ile Trp Thr Glu Gly Arg Met Lys Leu Asn Asp Ile Ile Thr Glu Asn 
                165                 170                 175 

Lys Asn Val Ser Asp Arg Ala Thr Ala Ala Ile Asp Glu Ala Val Leu 
            180                 185                 190 

Ser Ala Lys Ile Ser Met Ala Val Ser Leu Leu Ile Ala Ile Leu Ala 
        195                 200                 205 

Ala Gly Leu Cys Gly Leu Leu Leu Met Arg Ala Ile Met Ala Pro Met 
    210                 215                 220 

Gln Arg Ile Val Asp Ile Leu Glu Thr Met Arg Asp Gly Asp Leu Ser 
225                 230                 235                 240 

Lys Arg Leu Asn Leu Glu Arg Lys Asp Glu Phe Gly Ala Val Glu Thr 
                245                 250                 255 

Gly Phe Asn Asp Met Met Thr Glu Leu Thr Ala Leu Val Ser Gln Ala 
            260                 265                 270 

Gln Arg Ser Ser Val Gln Val Thr Thr Ser Val Thr Glu Ile Ala Ala 
        275                 280                 285 

Thr Ser Lys Gln Gln Gln Ala Thr Ala Thr Glu Thr Ala Ala Thr Thr 
    290                 295                 300 

Thr Glu Ile Gly Ala Thr Ser Arg Glu Ile Ala Ala Thr Ser Lys Asp 
305                 310                 315                 320 

Leu Val Arg Thr Met Thr Glu Val Ser Thr Ala Ala Asp Gln Ala Ser 
                325                 330                 335 

Val Ala Ala Gly Ser Gly Gln Gln Gly Leu Ala Arg Met Glu Glu Thr 
            340                 345                 350 

Met His Ser Val Met Gly Ala Ala Asp Leu Val Asn Ala Lys Leu Ala 
        355                 360                 365 

Ile Leu Asn Glu Lys Ala Gly Asn Ile Asn Gln Val Val Val Thr Ile 
    370                 375                 380 

Val Lys Val Ala Asp Gln Thr Asn Leu Leu Ser Leu Asn Ala Ala Ile 
385                 390                 395                 400 

Glu Ala Glu Lys Ala Gly Glu Tyr Gly Arg Gly Phe Ala Val Val Ala 
                405                 410                 415 

Thr Glu Val Arg Arg Leu Ala Asp Gln Thr Ala Val Ala Thr Tyr Asp 
            420                 425                 430 

Ile Glu Gln Met Val Arg Glu Ile Gln Ser Ala Val Ser Ala Gly Val 
        435                 440                 445 

Met Gly Met Asp Lys Phe Ser Glu Glu Val Arg Arg Gly Met Phe Glu 
    450                 455                 460 

Val Gln Gln Val Gly Glu Gln Leu Ser Gln Ile Ile His Gln Val Gln 
465                 470                 475                 480 

Ala Leu Ala Pro Arg Val Leu Met Val Asn Glu Gly Met Gln Ala Gln 
                485                 490                 495 

Ala Thr Gly Ala Glu Gln Ile Asn His Ala Leu Val Gln Leu Gly Asp 
            500                 505                 510 

Ala Ser Ser Gln Thr Val Glu Ser Leu Arg Gln Ala Ser Phe Ala Ile 
        515                 520                 525 

Asp Glu Leu Ser Gln Val Ala Val Gly Leu Arg Ser Gly Val Ser Arg 
    530                 535                 540 

Phe Lys Val 
545 

 
           
             29  
             170  
             PRT  
             Pseudomonas fluorescens  
           
            29 

Met Ser Glu Leu Ala Ala Lys Arg Gly Ala Val Pro Ala Ala Lys Lys 
  1               5                  10                  15 

Ala Leu Phe Leu Val Phe His Ile Gly Gln Glu Arg Tyr Ala Leu Lys 
             20                  25                  30 

Ala Thr Glu Val Ala Glu Val Leu Pro Arg Leu Pro Leu Lys Pro Ile 
         35                  40                  45 

Ala His Ala Pro Leu Trp Val Ala Gly Ile Phe Ala His Arg Gly Ala 
     50                  55                  60 

Leu Val Pro Val Ile Asp Leu Ser Ala Leu Thr Phe Gly Asn Pro Ala 
 65                  70                  75                  80 

Gln Ala Arg Thr Ser Thr Arg Leu Val Leu Val Asn Tyr Gln Pro Asp 
                 85                  90                  95 

Ala Gly Ser Gln Ala Arg Trp Leu Gly Leu Ile Leu Glu Gln Ala Thr 
            100                 105                 110 

Asp Thr Leu Arg Cys Asp Pro Ala Glu Phe Gln Pro Tyr Gly Leu Ala 
        115                 120                 125 

Asn Arg Gln Ala Pro Tyr Leu Gly Pro Val Arg Glu Asp Ala Leu Gly 
    130                 135                 140 

Leu Met Gln Trp Ile Gly Val Asn Asp Leu Leu Thr Asp Asp Val Arg 
145                 150                 155                 160 

Ala Val Leu Phe Ser Ala Glu Leu Ser Val 
                165                 170 

 
           
             30  
             419  
             PRT  
             Pseudomonas fluorescens  
           
            30 

Met Ser Asn Asp Pro Arg Phe Phe Ala Phe Leu Lys Glu Arg Ile Gly 
  1               5                  10                  15 

Leu Asp Val Ala Ser Val Gly Glu Ala Ile Ile Glu Arg Ala Val Arg 
             20                  25                  30 

Gln Arg Ser Gln Ile Val Gln Ala Pro Thr Pro Gly Glu Tyr Trp Gln 
         35                  40                  45 

His Leu Gln Ser Ser Gln Asp Glu Gln Gln Ala Leu Ile Glu Ala Val 
     50                  55                  60 

Ile Val Pro Glu Thr Trp Phe Phe Arg Tyr Pro Glu Ser Phe Ala Thr 
 65                  70                  75                  80 

Leu Ala Arg Leu Ala Lys Ala Arg Leu Val Asp Ile Lys Gln Met Arg 
                 85                  90                  95 

Ala Leu Arg Ile Leu Ser Leu Pro Cys Ser Thr Gly Glu Glu Pro Tyr 
            100                 105                 110 

Ser Ile Ala Met Ala Leu Leu Asp Ala Gly Leu Ala Pro His Gln Phe 
        115                 120                 125 

Lys Val Gln Gly Met Asp Val Ser Pro Leu Ser Val Glu Arg Ala Arg 
    130                 135                 140 

Arg Gly Val Tyr Gly Lys Asn Ser Phe Arg Gly Gly Asp Ile Ala Phe 
145                 150                 155                 160 

Arg Asp Arg His Phe Thr Glu Tyr Gly Asp Gly Phe His Ile Ala Asp 
                165                 170                 175 

Arg Val Arg Glu Gln Val Arg Leu Gln Val Gly Asn Leu Leu Asp Pro 
            180                 185                 190 

Ala Leu Leu Val Asn Glu Ala Ala Tyr Asp Phe Val Phe Cys Arg Asn 
        195                 200                 205 

Leu Leu Ile Tyr Phe Asp Gln Pro Thr Gln Lys Gln Val Phe Asp Val 
    210                 215                 220 

Leu Lys Gly Leu Thr His Val Asp Gly Val Leu Phe Ile Gly Pro Ala 
225                 230                 235                 240 

Glu Gly Ser Leu Leu Gly Arg His Gly Met Arg Ser Ile Gly Val Pro 
                245                 250                 255 

Gln Ser Phe Ala Phe Ser Arg Gln Gly Ser Pro Gln Leu Pro Glu Pro 
            260                 265                 270 

Ala Phe Ile Pro Thr Pro Ala Pro Thr Pro Pro Arg Ser Thr Ala Pro 
        275                 280                 285 

Ile Ser Ala Lys Pro Arg Pro Phe Ser Thr Val Ser Ala His Val Leu 
    290                 295                 300 

Pro Ile Lys Ala Thr Pro Ser Asp Ala Gly Thr Leu Leu Ser Arg Ile 
305                 310                 315                 320 

Ala Thr Leu Ala Asn Glu Gly Lys Ser Ala Glu Ala Arg Ala Ala Cys 
                325                 330                 335 

Glu Asp Tyr Leu Asn Ser His Pro Pro Ala Ala Gln Val Phe Tyr Trp 
            340                 345                 350 

Leu Gly Leu Leu Ser Asp Val Ala Gly Ser Ala Leu Glu Ala Gln Gly 
        355                 360                 365 

Tyr Tyr Arg Lys Ala Leu Tyr Leu Glu Pro Gln His Pro Gln Ala Leu 
    370                 375                 380 

Met His Leu Ala Ala Leu Leu Glu Ser Arg Gly Asp Ser Ala Gly Ala 
385                 390                 395                 400 

Arg Arg Leu Gln Ala Arg Ala Ala Arg Ser Glu Arg Ala Asp Ser Glu 
                405                 410                 415 

Ser Lys Pro 

 
           
             31  
             232  
             PRT  
             Pseudomonas fluorescens  
           
            31 

Met Ser Asn Thr Glu Ala Leu Asp Thr Thr Gly Leu Asp Leu Thr Leu 
  1               5                  10                  15 

Ala Asp Thr Gln Ala Ile Asp Asp Cys Trp Asn Arg Ile Gly Ile His 
             20                  25                  30 

Gly Asp Lys Ser Cys Pro Leu Leu Ala Asp His Ile His Cys Arg Asn 
         35                  40                  45 

Cys Ser Val Tyr Ser Ala Ala Ala Thr Arg Leu Leu Asp Arg Tyr Ala 
     50                  55                  60 

Leu Gln Gln Asp Asp Arg Arg Pro Gln Ala Ala Glu Val Asp Thr Glu 
 65                  70                  75                  80 

Val Val Thr Arg Ser Leu Leu Met Phe Arg Leu Gly Glu Glu Trp Leu 
                 85                  90                  95 

Gly Ile Ala Thr Arg Cys Leu Val Glu Val Ala Pro Leu Gln Pro Ile 
            100                 105                 110 

His Ser Leu Pro His Gln Arg Ser Arg Ala Leu Leu Gly Val Ala Asn 
        115                 120                 125 

Val Arg Gly Ala Leu Val Ala Cys Leu Ser Leu Val Glu Leu Leu Gly 
    130                 135                 140 

Leu Asp Ala Thr Ser Ser Gly Ala Thr Gly Gly Arg Ile Met Pro Arg 
145                 150                 155                 160 

Met Leu Ile Ile Ala Ala Gln Asp Gly Pro Val Val Val Pro Val Asp 
                165                 170                 175 

Glu Val Asp Gly Ile His Ala Ile Asp Glu Arg Thr Leu Lys Ala Ala 
            180                 185                 190 

Ser Ala Ser Gly Thr Gln Ala Ser Ala Arg Phe Thr Gln Gly Val Leu 
        195                 200                 205 

Pro Trp Lys Gly Arg Ser Leu Arg Trp Leu Asp Glu Ala Gln Leu Leu 
    210                 215                 220 

Ser Ala Val Thr Arg Ser Leu Ser 
225                 230 

 
           
             32  
             755  
             PRT  
             Pseudomonas fluorescens  
           
            32 

Met Thr Pro Asp Gln Met Arg Asp Ala Ser Leu Leu Glu Leu Phe Ser 
  1               5                  10                  15 

Leu Glu Ala Asp Ala Gln Thr Gln Val Leu Ser Ala Gly Leu Leu Ala 
             20                  25                  30 

Leu Glu Arg Asn Pro Thr Gln Ala Asp Gln Leu Glu Ala Cys Met Arg 
         35                  40                  45 

Ala Ala His Ser Leu Lys Gly Ala Ala Arg Ile Val Gly Val Asp Ala 
     50                  55                  60 

Gly Val Ser Val Ser His Val Met Glu Asp Cys Leu Val Ser Ala Gln 
 65                  70                  75                  80 

Glu Asn Arg Leu Tyr Leu Gln Pro Glu His Ile Asp Ala Leu Leu Gln 
                 85                  90                  95 

Gly Thr Asp Leu Leu Met Arg Ile Ala Thr Pro Gly Asn Asp Val Gly 
            100                 105                 110 

Pro Ala Asp Val Glu Ala Tyr Val Ala Leu Met Glu Arg Leu Leu Asp 
        115                 120                 125 

Pro Ser Gln Ala Pro Val Asn Val Ala Pro Ser Pro Glu Pro Ala Pro 
    130                 135                 140 

Val Val Glu Glu Leu Pro Pro Glu Pro Glu Pro Ala Pro Pro Val Asn 
145                 150                 155                 160 

Ser Glu Pro Pro Arg Gln Gly Lys Arg Met Thr Glu Gly Gly Glu Arg 
                165                 170                 175 

Val Leu Arg Val Thr Ala Glu Arg Leu Asn Ser Leu Leu Asp Leu Ser 
            180                 185                 190 

Ser Lys Ser Leu Val Glu Thr Gln Arg Leu Lys Pro Tyr Leu Ala Ser 
        195                 200                 205 

Leu Gln Arg Leu Lys Arg Ile Gln Ser Gln Gly Ile Arg Ala Leu Asp 
    210                 215                 220 

Thr Leu Asp Gly Gln Leu Lys Thr Gln Val Leu Ser Leu Glu Ala Gln 
225                 230                 235                 240 

Glu Ala Leu Ala Asp Thr Arg Arg Leu Leu Ser Glu Ala Gln Ala Leu 
                245                 250                 255 

Leu Ala Glu Lys His Ala Glu Leu Asp Glu Phe Gly Trp Gln Ala Gly 
            260                 265                 270 

Gln Arg Ala Gln Val Leu Tyr Asp Thr Ala Leu Ala Cys Arg Met Arg 
        275                 280                 285 

Pro Phe Ala Asp Val Leu Ala Gly Gln Val Arg Met Val Arg Asp Leu 
    290                 295                 300 

Gly Arg Ser Leu Gly Lys Gln Val Arg Leu Glu Ile Glu Gly Glu Lys 
305                 310                 315                 320 

Thr Gln Val Asp Arg Asp Val Leu Glu Lys Leu Glu Ala Pro Leu Thr 
                325                 330                 335 

His Leu Leu Arg Asn Ala Val Asp His Gly Ile Glu Met Pro Glu Gln 
            340                 345                 350 

Arg Leu Leu Ala Gly Lys Pro Ala Glu Gly Leu Ile Arg Leu Arg Ala 
        355                 360                 365 

Ser His Gln Ala Gly Leu Leu Val Leu Glu Leu Ser Asp Asp Gly Asn 
    370                 375                 380 

Gly Val Asp Leu Glu Arg Leu Arg Gly Thr Ile Val Asp Arg His Leu 
385                 390                 395                 400 

Ser Pro Val Glu Thr Ala Leu Arg Leu Ser Glu Glu Glu Leu Leu Thr 
                405                 410                 415 

Phe Leu Phe Leu Pro Gly Phe Ser Leu Arg Asp Thr Val Thr Glu Val 
            420                 425                 430 

Ser Gly Arg Gly Val Gly Leu Asp Ala Val Gln His Met Val Arg Gln 
        435                 440                 445 

Leu Arg Gly Ala Val Val Leu Glu Gln Thr Ala Gly Gln Gly Ser Arg 
    450                 455                 460 

Phe His Leu Glu Val Pro Leu Thr Leu Ser Val Val Arg Ser Leu Val 
465                 470                 475                 480 

Val Glu Val Gly Glu Glu Ala Tyr Ala Phe Pro Leu Ala His Ile Glu 
                485                 490                 495 

Arg Met Cys Asp Leu Ala Pro Asp Asp Ile Val Gln Leu Glu Gly Arg 
            500                 505                 510 

Gln His Phe Trp His Glu Gly Arg His Val Gly Leu Val Ala Ala Ser 
        515                 520                 525 

Gln Leu Leu Gln Arg Pro Ala Gly Gln Ser Pro Ser Glu Thr Leu Lys 
    530                 535                 540 

Val Val Val Ile Arg Glu Arg Asp Thr Val Tyr Gly Ile Ala Val Glu 
545                 550                 555                 560 

Arg Phe Ile Gly Glu Arg Thr Leu Val Val Leu Pro Leu Asp Asp Arg 
                565                 570                 575 

Leu Gly Lys Val Gln Asp Ile Ser Ala Gly Ala Leu Leu Asp Asp Gly 
            580                 585                 590 

Ser Val Val Leu Ile Val Asp Val Glu Asp Met Leu Arg Ser Val Asp 
        595                 600                 605 

Lys Leu Leu Asn Thr Gly Arg Leu Glu Arg Ile Ala Arg Arg Ser Gln 
    610                 615                 620 

Gln Thr Thr Glu Ala Pro Arg Lys Arg Val Leu Val Val Asp Asp Ser 
625                 630                 635                 640 

Leu Thr Val Arg Glu Leu Gln Arg Lys Leu Leu Leu Asn Arg Gly Tyr 
                645                 650                 655 

Glu Val Ala Val Ala Val Asp Gly Met Asp Gly Trp Asn Ala Leu Arg 
            660                 665                 670 

Ser Glu Asp Phe Asp Leu Leu Ile Thr Asp Ile Asp Met Pro Arg Met 
        675                 680                 685 

Asp Gly Ile Glu Leu Val Thr Leu Leu Arg Arg Asp Ser Arg Leu Gln 
    690                 695                 700 

Ser Leu Pro Val Met Val Val Ser Tyr Lys Asp Arg Glu Glu Asp Arg 
705                 710                 715                 720 

Arg Arg Gly Leu Asp Ala Gly Ala Asp Tyr Tyr Leu Ala Lys Ala Ser 
                725                 730                 735 

Phe His Asp Asp Ala Leu Leu Asp Ala Val Val Glu Leu Ile Gly Gly 
            740                 745                 750 

Ala Arg Ala 
        755 

 
           
             33  
             336  
             PRT  
             Pseudomonas fluorescens  
           
            33 

Met Arg Ile Ala Ile Val Asn Asp Met Pro Leu Ala Val Glu Ala Leu 
  1               5                  10                  15 

Arg Arg Ala Leu Ser Phe Glu Pro Ala His Gln Val Val Trp Val Ala 
             20                  25                  30 

Ser Asn Gly Leu Glu Ala Val Gln Arg Cys Ala Glu Leu Thr Pro Asp 
         35                  40                  45 

Leu Ile Leu Met Asp Leu Ile Met Pro Val Met Asp Gly Val Glu Ala 
     50                  55                  60 

Thr Arg Gln Ile Met Ala Glu Thr Pro Cys Ala Ile Val Ile Val Thr 
 65                  70                  75                  80 

Val Asp Arg Gln Ala Asn Val Ser Arg Val Phe Glu Ala Met Gly His 
                 85                  90                  95 

Gly Ala Leu Asp Val Val Asp Thr Pro Pro Leu Gly Val Gly Asn Pro 
            100                 105                 110 

Lys Asp Ala Ala Ala Pro Leu Leu Arg Lys Ile Leu Asn Ile Gly Trp 
        115                 120                 125 

Leu Ile Gly Gln Arg Gly Thr Arg Val Arg Ala Glu Thr Leu Pro Ala 
    130                 135                 140 

Arg Ala Ser Gly Lys Arg Gln Ser Leu Val Ala Ile Gly Ser Ser Ala 
145                 150                 155                 160 

Gly Gly Pro Ala Ala Leu Glu Ile Leu Leu Lys Gly Leu Pro Arg Asp 
                165                 170                 175 

Phe Pro Ala Ala Ile Val Leu Val Gln His Val Asp Gln Val Phe Ala 
            180                 185                 190 

Ala Gly Met Ala Glu Trp Leu Ser Ser Ala Ser Gly Leu Pro Val Arg 
        195                 200                 205 

Leu Ala Arg Glu Gly Glu Pro Pro Gln Ser Gly Val Val Leu Leu Ala 
    210                 215                 220 

Gly Thr Asn His His Ile Arg Leu Leu Lys Asn Gly Thr Leu Ala Tyr 
225                 230                 235                 240 

Thr Ala Glu Pro Val Asn Glu Ile Tyr Arg Pro Ser Ile Asp Val Phe 
                245                 250                 255 

Phe Glu Ser Val Ala Ser His Trp Asn Gly Asp Ala Val Gly Val Leu 
            260                 265                 270 

Leu Thr Gly Met Gly Arg Asp Gly Ala Gln Gly Leu Lys Leu Leu Arg 
        275                 280                 285 

Glu Gln Gly Tyr Leu Thr Ile Ala Gln Asp Gln Gln Ser Ser Ala Val 
    290                 295                 300 

Tyr Gly Met Pro Lys Ala Ala Ala Ala Ile Asp Ala Ala Val Glu Ile 
305                 310                 315                 320 

Arg Pro Leu Asp Arg Ile Ala Pro Arg Leu Leu Glu Val Phe Ala Lys 
                325                 330                 335 

 
           
             34  
             333  
             PRT  
             Pseudomonas fluorescens  
           
            34 

Met Asn Asp Leu Gln Ile Asp Asp Ile Lys Thr Asp Glu Asn Ala Ala 
  1               5                  10                  15 

Met Val Leu Leu Val Asp Asp Gln Ala Met Ile Gly Glu Ala Val Arg 
             20                  25                  30 

Arg Gly Leu Ala His Glu Glu Asn Ile Asp Phe His Phe Cys Ala Asp 
         35                  40                  45 

Pro His Gln Ala Ile Ala Gln Ala Ile Arg Ile Lys Pro Thr Val Ile 
     50                  55                  60 

Leu Gln Asp Leu Val Met Pro Gly Leu Asp Gly Leu Thr Leu Val Arg 
 65                  70                  75                  80 

Glu Tyr Arg Asn His Pro Ala Thr Gln Asn Ile Pro Ile Ile Val Leu 
                 85                  90                  95 

Ser Thr Lys Glu Asp Pro Leu Ile Lys Ser Ala Ala Phe Ser Ala Gly 
            100                 105                 110 

Ala Asn Asp Tyr Leu Val Lys Leu Pro Asp Asn Ile Glu Leu Val Ala 
        115                 120                 125 

Arg Ile Arg Tyr His Ser Arg Ser Tyr Met Thr Leu Leu Gln Arg Asp 
    130                 135                 140 

Ala Ala Tyr Arg Ala Leu Arg Val Ser Gln Gln Gln Leu Leu Asp Thr 
145                 150                 155                 160 

Asn Leu Val Leu Gln Arg Leu Met Asn Ser Asp Gly Leu Thr Gly Leu 
                165                 170                 175 

Ser Asn Arg Arg His Phe Asp Glu Tyr Leu Glu Leu Glu Trp Arg Arg 
            180                 185                 190 

Ala Met Arg Asp Gln Thr Gln Leu Ser Leu Leu Met Ile Asp Val Asp 
        195                 200                 205 

Phe Phe Lys Thr Tyr Asn Asp Ser Phe Gly His Val Glu Gly Asp Glu 
    210                 215                 220 

Ala Leu Arg Lys Val Ala Ala Thr Ile Arg Glu Ala Ser Ser Arg Pro 
225                 230                 235                 240 

Ser Asp Leu Pro Ala Arg Tyr Gly Gly Glu Glu Phe Ala Leu Val Leu 
                245                 250                 255 

Pro Asn Thr Ser Pro Gly Gly Ala Arg Leu Val Ala Glu Lys Leu Arg 
            260                 265                 270 

Met Ala Val Ala Ala Leu Lys Ile Pro His Ile Ala Pro Thr Glu Gly 
        275                 280                 285 

Ser Ser Leu Thr Ile Ser Ile Gly Leu Ser Thr Met Thr Pro Gln Gln 
    290                 295                 300 

Gly Thr Asp Cys Arg Gln Val Ile Val Ala Ala Asp Lys Gly Leu Tyr 
305                 310                 315                 320 

Thr Ala Lys His Asn Gly Arg Asn Gln Val Gly Ile Glu 
                325                 330