NUCLEIC ACID PROBES FOR DETECTING E. COLI O157:H7

Abstract: An isolated nucleic acid molecule that hybridizes under stringent conditions to SEQ ID NO:1 or its complement and to the DNA of enterohemorrhagic E. coli O157:H7 but not to the DNA of enteropathogenic E. coli O55:H7.

Description: NUCLEIC ACID PROBES FOR DETECTING E. coli 0157:H7 Field of the Invention The invention relates to genetic engineering and provides nucleic acid probes for detecting pathogenic Escherichia coli O157:H7 in food and fecal samples. Background of the Invention E. coli O157:H7 is a virulent food-borne pathogen that causes acute hemorrhagic colitis (bloody inflammation of the colon), and sometimes a severe hemolytic uremic syndrome (HUS), in children. These conditions can be fatal, or cause permanent kidney damage and shortened life expectancy. Treatment options for this infection are limited; prevention of human infection is therefore of the greatest importance. Epidemics of E. coli O157:H7 generally have been traced to contaminated water or meats, especially undercooked hamburger. Following the 1993 massive outbreak in Washington State, the strain responsible for the epidemic disappeared soon after the recall of the incriminated vehicle. Hence, ingestion of contaminated beef, and not person to person spread, appears to be the chief source of human infection. Because of the rapid onset of life-threatening complications, expedient diagnosis is of utmost importance to those infected with this strain of E. coli, especially when the patient is a child. However, because the early symptoms of this disease can mimic other disorders, patients infected with this organism during the initial stages of new epidemics may not be tested for E. coli O157:H7 until other causes have been ruled out. Once E. coli 0\ 57.H7 is suspected, diagnostic testing methods currently available require about a day to complete before a definitive diagnosis can be confirmed. Any means for shortening this dangerous delay would reduce the risk to infected patients and family members who might become infected through person to person transmission. Of utmost importance for controlling outbreaks of E. coli 0157 :H7 is the rapid detection of the contaminated food source. A common method for detecting E. coli 0157 :H7 includes incubation on agar containing sorbitol, a substrate that supports growth of most fecal E. coli strains, but which the O157:H7 strain cannot metabolize. However, failure to ferment sorbitol does not provide a definitive diagnosis, as non-pathogenic strains of E. coli exist that also cannot ferment sorbitol. A latex agglutination test for identification of the serogroup 0157 is available from Oxoid (Unipath Limited, Basingstoke, Hampshire, England), which recommends that the test be applied to isolates that already have been determined to lack the ability to ferment sorbitol. This antibody-based test lacks specificity: Some strains of E. hermanii share the antigen detected by the antiserum; hence this test must be confirmed by further fermentation testing before an isolate can be conclusively identified as E. coli 0157:H7. Furthermore, noncytotoxic E. coli may possess the E. coli 0157 antigen, leading to false positive reactions in such 0157 antigen detection systems. Other diagnostic methods for E. coli 0157 H7 have been suggested. Immunofluorescent examination of fresh stool with labeled 0157-specific antiserum has recently been proposed as a rapid technique for the detection of patients whose stools contain E. coli 0157 :H7 (Park et al., Am. J. Clin. Path. 101:91-94, 1994). Antibody-coated magnetic beads (Dynal) are available that react with E. coli O157:H7 However, antigen detection tests should be accompanied by a confirmatory stool culture (Tarr, Clinical Infectious Diseases, 20: 1-8, 1995). A test kit for E. coli O157:H7 is available which reportedly can detect this organism in meat (Organon Teknika, Durham, NC). This is an antigen-based technology, in which bacterial antigens are used as targets to identify the organism. The test requires a day (i.e., overnight) to produce a presumptive positive test. An additional technique which has been proposed is based on a rapid dipstick immunoassay to detect enterohemorrhagic E. coli O157:H7 in retail ground beef (Kim et al. , Applied Environmental Micobiology 58:1764- 1767, 1992). For diagnostic purposes, conventional microbiological and antibody-based testing techniques may be insufficient for management of epidemic outbursts of HUS. By the time HUS appears, about two-thirds of patients no longer have E. coli O157:H7 in their stools. Moreover, the numbers of organisms present in contaminated food samples often are too low to be readily detected. DNA probes specific for this pathogen would provide an alternative method that is capable of detecting small numbers of organisms. Most efforts in the past to develop such probes have focused on the gene encoding the Shiga-lϊke toxins (SLTs) believed to be responsible for many of strain O157's pathogenic effects. See: Levine et al., The Journal of Infectious Diseases 156(1):175-182, 1987; Samadpour et al., Applied and Environmental Microbiology 56(5):1212-1215, 1990; Pollard et al., Journal of Clinical Microbiology 28(3):540-545, 1990; Pollard et al., The Journal of Infectious Diseases 162:1195-1198, 1990. However, the pathogenecity of many strains containing SLT genes is questionable (Tarr, Clinical Infectious Diseases, 20:1-8, 1995). PCR assays to detect O157:H7 strains using the uidA gene have also been reported (Feng et al., Applied and Environmental Microbiology 57(1): 320-323, 1991; Feng, Molecular and Cellular Probes 7:151-154, 1993.) More recently, a multiplex PCR assay was reported which uses three sets of primers, two of which are directed to conserved regions within genes encoding for SLT-I and SLT-II, and the third set directed to the uidA gene (Cebula et al., Journal of Clinical Microbiology 33(l):248-250, 1995). Summary of the Invention The invention provides a nucleic acid probe for detecting the presence of enterohemorrhagic E. coli 0157. 7, comprising an isolated nucleic acid molecule that hybridizes under stringent conditions to SEQ ID NO:l or its complement and to the DNA of E. coli O157:H7 but not to the DNA of the clonally related enteropathogenic E. coli 055 :H7. Detailed Description of the Preferred Embodiment The invention provides a DNA fragment (SEQ ED NO:l) that is useful for detecting E. coli O157:H7 in food and fecal samples. This DNA fragment, isolated from E. coli O157:H7, does not cross-hybridize with the known genes for Shiga toxins in Shigella dysenteriae, or the uidA gene which encodes for β-glucoronidase in E. coli. Rather, this DNA fragment unexpectedly contains two genes that are related by homology to the putative transmembrane export protein of Yersinia pseudotuberculosis (Kessler et al., Journal of Bacteriology 175(5): 1412-1422, 1993), and to the rflE gene which reportedly encodes perosamine synthetase in Vibrio cholerae (Stroeher et al., Proc Natl. Acad. Sci. USA 89:2566-2570, 1992). The subject DNA fragment was serendipitously identified through efforts to determine the role of the 0157 lipopolysaccharide in the pathogenesis . of E. coli O157:H7. Briefly summarized, IxφhoA insertion mutagenesis was used to create mutants of£. coli O157:H7 that did not express this antigen. It was noted that such mutants were hyperadherent, that is, they had a pronounced increase in their ability to adhere to HeLa cells. Two 0157 * hyperadherent mutants, designated 20D2B and F12, were selected for detailed analysis after screening ca. 3000 TiφhoA mutants of E. coli O157:H7 for the loss of the antigen by the latex particle agglutination test. Both mutant strains were approximately eight-fold more adherent to HeLa cells than the parent strain. Southern blots indicated that these mutants contained two and one TrφhoA insertions, respectively. Nucleotide sequence analysis of the DNA regions flanking the F 12 insertion surprisingly revealed two open reading frames that share significant homology with the putative transmembrane export protein of Yersinia pseudotuberculosis and the rβE gene of Vibrio cholerae. Using PCR primers (identified below) from these flanking regions, the subject DNA fragment was cloned from E. coli O157:H7. This fragment is referred to herein as "the F12 region" and its nucleotide sequence is depicted in SEQ LD NO:l. The invention provides nucleic acid probes derived from the F12 region. Preferred probes are selected from the two open reading frames corresponding to nucleotides 1-1026 (SEQ LD NO:2) and nucleotides 1029-2123 (SEQ ID NO:3) shown in SEQ LD NO:l. By way of example, use of a nucleic acid probe (SEQ LD NO:4), corresponding to nucleotides 913-2199 shown in SEQ ID NO:l, is described below. Other nucleic acid probes can be readily selected from among the contiguous nucleotides within the disclosed sequence(s) and screened for both sensitivity (for strain O157:H7) and specificity (lack of sensitivity for other microbial contaminants of food and feces) using an appropriate panel of positive and negative genomic controls. In the screening assay, E. coli O157:H7 serves as the positive genomic control. Negative genomic controls are selected from among other microbial strains that typically contaminate the type of food, agricultural, or clinical sample to be tested. Such negative genomic controls are preferably selected from among E. coli strains that are genetically and antigenically related and most preferably clonally related to strain O157:H7. The clonal relationships among E. coli strains that cause hemorrhagic colitis and infantile diarrhea are described in Whittam et al., Infection and Immunity 61:1619-1629, 1993. Figure 4 therein presents a phylogenetic tree of genomically closely related strains. In the same lineage as strain O157:H7 are the following clonally related strains which can serve as suitable negative controls, notably: the enteropathogenic strains 055 :H7 (i.e., the closest phylogenetically related strain), Ol l l:H12, and O55:H6. Other E. coli strains suitable as negative controls include noncytotoxic and cytotoxic E. coli of serotypes O157:H43, Ol l l:H8, O26:Hl l, O128:H7, O128.H21, Ol l l:H21, O128:H2, and Ol l l:H2. Bacterial strains for use in the screening panel can be obtained from the reference collections of: the Centers for Disease Control (CDC), Atlanta, GA; the E. coli Reference Center, University Park, PA; the American Type Culture Collection (ATCC), Rockville, MD; the Center for Vaccine Development, Baltimore, MD; the Division of Microbiology, Food and Drug Administration (FDA), Washington, D.C.; the Center for Disease Control, Ottawa, Canada; the Universitats-Krankenhaus, Hamburg, Germany; and the Statens Seruminstitut, Copenhagen, Denmark. Such a reference panel of bacteria can be employed to identify nucleic acid probes from the F12 region shown in SEQ ID NO: l that provide the requisite sensitivity and selectivity for distinguishing E. coli O157:H7 from other bacteria, including related strains of E. coli. In a representative screening system, bacteria in the test panel are plated on trypticase soy agar, as described in Samadpour et al., 1990, which is hereby incorporated by reference. Candidate oligonucleotide probes randomly chosen from contiguous nucleotides within the F12 region are synthesized using a DNA synthesizer, following the procedures recommended by the manufacturer. A probe corresponding to SEQ LD NO:4, which has been shown to possess the requisite specificity, may be used as a control probe. All probes are labeled using conventional techniques. Aliquots of chromosomal DNA extracted from each strain of bacteria in the panel are hybridized under stringent conditions with the labeled candidate and control probes. Wild-type E. coli O157:H7 serves as the positive control. Candidate probes that hybridize under stringent conditions with O157:H7 but only slightly or not at all with the negative control strains are deemed to have the requisite specificity for detecting E. coli O157:H7 in samples of ground meat, bovine feces, and samples of clinical interest. The F12 fragment (SEQ ID NO:l) can also be employed to identify nucleic acid probes from the E. coli O157:H7 genome flanking the F12 region that provide the requisite sensitivity and selectivity. To identify these contiguous sequences using the F12 region, a library of random DNA fragments is generated by partial endonuclease digestion, cloned into an appropriate vector, and screened with the F12 probe. Cloned DNA is mapped to determine if regions contiguous to the F12 region are present on the inserted DNA identified by homology to the F12 region. The contiguous DNA is sequenced, and tested for specificity and sensitivity as a probe for the detection of£. coli O157:H7. The nucleotide length of the probe is chosen using customary criterion. Ten nucleotides is generally the lower limit for such nucleotide segments, because sequences smaller than that sometimes fail to form stable hybrid duplexes, or may hybridize nonspecifically with unrelated genes that happen to share short stretches of complementary sequence with the desired target sequence. Accordingly, in preferred embodiments, the probes are at least 10 nucleotides in length, with the oligσnucleotide being capable of forming a detectable stable duplex with the F12 region sequence(s). More preferred are probes of at least 15 nucleotides, as such probes are more likely to form stable duplexes with their complementary sequences. Oligonucleotides at least 15 bases long are very likely to be specific, as sequences of this length will occur by chance no more than once in a mammalian genome, hence are extremely unlikely to occur by chance in a bacterial genome. Preferred probes will range from 15 to 50 bases in length, as probes in this range can be prepared at a reasonable cost using commercially available DNA synthesizers or services that synthesize oligonucleotides on request. The invention also contemplates that fragments of DNA even longer than 50 nucleotides may be obtained from the F12 region, by using restriction endonucleases, or by using PCR primers and subsequently cloning these longer fragments. For use in the described selection system or for analyzing food or fecal samples, probes must be labeled in order to be detectable in hybrids formed during the assays. Synthetic probes are conventionally labeled with radioactive phosphorus using polynucleotide kinase. Alternatively the probes can be tagged with an affinity reagent such as biotin or streptavidin, thus enabling the probe to bind to an enzyme capable of cleaving a chromogenic substrate (for details, see Sambrook et al.). Various conventional protocols can be employed for the hybridization step. Generally, the labeled probe is present in a suitable hybridization solution, while the target nucleic acids are fixed to an insoluble substratum, such as derivatized nylon. In the preferred embodiment, target DNA sequences and labeled probes are incubated under stringent hybridization conditions. Such conditions are generally understood to be those that permit only perfectly matched or nearly perfectly matched hybrids to form, and their use will assure that probes hybridize specifically with sequences of E. coli O157:H7. The destabilizing effects of mismatched bases increases as probe length decreases, such that for probes shorter than 20 nucleotides, stringent hybridization conditions will rarely tolerate any mismatched bases. For longer probes, it is possible that a few mismatches may be present in hybrids that form even under stringent conditions. In some situations, practitioners could be misled by results involving mismatched hybrids, but in the present instance, the screening methods provided ensure identification of probes having the appropriate specificity regardless of whether mismatches may be present. That is, the screening procedure identifies probes that react with the targeted O157:H7 strain (and, optimally, with the positive control F12 region) but not with the negative control bacteria. Hence it is immaterial whether any given probe forms a perfect or only a near-perfect hybrid, as the ability to distinguish these two groups of bacteria is the only pertinent quality in the context of the present test assays. A variety of operable hybridization conditions are available to one skilled in the art, e.g., as provided in Sambrook et al. For probes longer than about 200 nucleotides in length, hybridization usually is carried out at about 25°C below the melting temperature of the hybrid, as this gives the optimal rate of duplex formation. For shorter probes, hybridization is usually carried out closer to the calculated melting temperature of a perfect hybrid, e.g., 10-15°C below the melting temperature. For probes of various lengths, melting temperatures for perfectly matched hybrids can be determined empirically, or can be calculated using one of the formulas found in Sambrook et al. An exemplary hybridization solution for oligonucleotides contains 6 X SSC (ca. 1 M [Na+]), and 0.5% sodium dodecyl sulfate. The subject sequences can also be used for detection of E. coli O157:H7 in polymicrobial samples using polymerase chain reaction (PCR) methodology. For example, nucleotide primers which have a high likelihood of being specific for E. coli O157:H7 are determined, using techniques described above, and used in PCR assays to detect genomic sequences specific for E. coli O157:H7 in enrichment cultures likely to contain the target organism. For example, a sample suspected of having E. coli O157:H7 in its natural condition (e.g., ground meat or bovine feces) is incubated in a broth enrichment culture, DNA is extracted using standard techniques, the DNA is annealed to the appropriate primers, in the PCR reaction, and the resulting bands are examined on agarose gels for products of this reaction. A positive band suggests the presence of £. coli O157:H7. It is estimated that such a technique could be performed within several hours of the start of incubation, that is, in a time period considerably less than present techniques which rely on amplification of the organism to an extent that antigens can be detected. In another embodiment, the expression products (SEQ LDS NO:5 and NO:6, respectively) of the nucleotide sequences shown in SEQ LO NOS: 2 and 3 are considered candidate immunogens for preparing antibody reagents for detection of the O157:H7 strain. The invention is further illustrated by the following example, which describes detection of E. coli 0157:117 using a nucleic acid probe (SEQ ED NO:4) encompassing the disclosed rβE homologous sequence (SEQ ID NO:3). EXAMPLE Total bacterial DNA (chromosome and plasmid) was prepared from a variety of bacteria, including: E. coli 0157 :rl7 strain 86-24, a strain which caused an outbreak of infection in a fast food restaurant in Walla Walla, Washington, in 1986 (Griffin et al., Annals of Internal Medicine, 109:705-712, 1988) as the positive control (the F12 region was derived from this strain); E. coli NM544 (a laboratory strain) as the negative control (Raleigh et al., Nucleic Acids Research, 16:1563-1575, 1988); five test strains from diarrheal E. coli group 5 (all E. coli 055 :H7); a clone of E. coli which is closely related to E. coli O157:H7 (Whittam et al., Infection and Immunity, 61:1619-1629, 1993); and five test strains of E. coli O157:H43. The DNA from each of these organisms was digested with EcoRI, separated electrophoretically in 0.7% agarose, and transferred to a nylon support membrane. The immobilized DNA was probed with a fragment of F12 DNA (SEQ ID NO:4), and homologous DNA sequences were detected in: E. coli 0157 :H7 strain 86-24; and four of the five E. coli O157.Η43 strains probed, though at lesser intensity. This probe (SEQ ED NO:4) identified an EcoRI fragment of ca. 6 kb in the strains which were positive. The probe did not identify homologous DNA in the negative control, the E. coli O55:H7 strains, and one of the E. coli O157:H43 strains. It is important to note, however, that the nonreactive E. coli O157:H43 strain was only very weakly positive in the Oxoid latex particle agglutination test (unlike the other E. coli O157.H43 strains), and its identity as a member of the 0157 serogroup is not certain. Plasmid pF12 containing SEQ ED NO:7 (i.e., nucleotides 32-2199 of SEQ ED NO:l) was deposited on April 14, 1995, at the American Type Culture Collection, 12301 Parklawn Drive, Rockville, MD 20852, U.S.A. pF12 is a PCR product of the cloned homologs of the V. cholerae rflE gene and the gene encoding the putative transmembrane export protein of Y. pseudotuberc losis from E. coli 0\ 57 : 7 strain 86-24, as well as some flanking sequence. It was cloned by using as primers the sequences 5'CTT CTG GCA TGA TTG ATT GGC3' (SEQ ID NO:8) and 3'CGA GTG GGG CGG TGG AAT TG5' (SEQ ED NO:9) from the deduced nucleotide sequences of the cloned DNA segments surrounding the TηpΛoA insertion in strain F 12. The primers were modified to contain BamHI and EcoRI sites, respectively, which were used to insert the PCR product into pSK+ (Stratagene) at these sites. Confirmatory sequencing from the PCR products and of the cloned regions flanking the TηpAoA insertion demonstrate two errors in these cloned segments. (SEQ ED NO: 1 is the correct sequence.) These include a T at position 964 (which is an erroneous C in the cloned insert deposited at ATCC) and a C at position 1510 (which is an erroneous T in the cloned insert deposited at ATCC). The practice of the present invention will employ, unless otherwise indicated, a number of conventional techniques of molecular and cellular biology, recombinant DNA, microbiology, and immunology, which are within the skill of the art. Such techniques are explained fully in the scientific literature and detailed experimental protocols are available in a number of technical manuals; see for example: F.M. Ausubel et al. (eds.), "Current Protocols in Molecular Biology," (1987 and 1993); Kriegler, M. (ed.), "Gene Transfer and Expression, a Laboratory Manual," (1990), W.H. Freeman Publishers; Sambrook, Fritsch, and Maniatis, "Molecular Cloning: A Laboratory Manual," Second Edition (1989); Erlich, H.A. (ed.) "PCR Technology: Principles and Applications for DNA Amplification," (1989), Stockton Press; Langone, J.J. and H. Van Vunakis (eds.), "Immunological Techniques, Part i: Hybridoma technology and monoclonal antibodies," Methods in Enzymology 121:1-947 (1986); Hurreil, J.G.R. (ed.), "Monoclonal Hybridoma Antibodies: Techniques and Applications," (1982), CRC Press, Boca Raton, FL; Coligan, J.E. et al. (eds), "Current Protocols in Immunology," (1991), Greene Publishing and Wiley- Interscience, N.Y.; Weir, D.M., "Handbook of Experimental Immunology," (1986), Blackwell Scientific; Kurstack, E., "Enzyme Immunodiagnostics," (1986), Academic Press, San Diego; Polak, J.M. and S. Van Noorden (eds.), "Immunocytochemistry: Practical Applications in Pathology and Biology," (1983), John Wright PSG, Littleton, MA; Sternberger, L.A., "Immunocytochemistry," (1986), Wiley, N.Y.; and primary references cited therein. All publications, patents, and patent publications mentioned herein are hereby incorporated herein by reference in their entirety. While the preferred embodiment of the invention has been illustrated and described, it will be appreciated that various changes can be made therein without departing from the spirit and scope of the invention. SEQUENCE LISTING (1) GENERAL INFORMATION: (i) APPLICANT: Tarr, Phillip I. Bilge, Si a S. Vary, Jr., James C. (ii) TITLE OF INVENTION: NUCLEIC ACID PROBES FOR DETECTING E. coli 0157:H7 (iii) NUMBER OF SEQUENCES: 9 (iv) CORRESPONDENCE ADDRESS: (A) ADDRESSEE: Christensen O'Connor Johnson & Kindness PLL (B) STREET: 1420 5th Ave., Suite 2800 (C) CITY: Seattle (D) STATE: WA (E) COUNTRY: USA (F) ZIP: 98101-2347 (v) COMPUTER READABLE FORM: (A) MEDIUM TYPE: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS (D) SOFTWARE: Patentln Release #1.0, Version #1.25 (vi) CURRENT APPLICATION DATA: (A) APPLICATION NUMBER: (B) FILING DATE: (C) CLASSIFICATION: (vii) PRIOR APPLICATION DATA: (A) APPLICATION NUMBER: US 08/423,564 (B) FILING DATE: 14-APR-1995 (viii) ATTORNEY/AGENT INFORMATION: (A) NAME: Shelton, Dennis K. (B) REGISTRATION NUMBER: 26,997 (C) REFERENCE/DOCKET NUMBER: CHOR-19417 (ix) TELECOMMUNICATION INFORMATION: (A) TELEPHONE: (206) 682-8100 (B) TELEFAX: (206) 224-0779 (2) INFORMATION FOR SEQ ID NO:l: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 2255 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (A) DESCRIPTION: nucleotide sequence of the F12 region (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Escherichia coli (xi) SEQUENCE DESCRIPTION: SEQ ID NO:l: TCGTTTGTCA TAACTGCAAT ATGCTATATT ACTTCTGGCA TGATTGATTG GCAACTAGTA 60 ATAAAAGGTA TAAACGAGAA TGTGTATGCA GAGTTACAAC ACTCAATTAA AGTCTTTGTA 120 ATCATATTTG GACTTGGAAT TTATTCAAAT GGTGTGCAAA AAGTTTATAT GGGAATACAA 180 AAAGCCTATA TAAGTAATAT TGTTAATGCC ATATTTATAT TGTTATCTAT TATTACTCTA 240 GTAATATCGT CGAAACTACA TGCGGGACTA CCAGTTTTAA TTGTCAGCAC TCTTGGTATT 300 CAATACATAT CGGGAATCTA TTTAACAATT AATCTTATTA TAAAGCGATT AATAAAGTTT 360 ACAAAAGTTA ACATACATGC TAAAAGAGAA GCTCCATATT TGATATTAAA CGGTTTTTTC 420 TTTTTTATTT TACAGTTAGG CACTCTGGCA ACATGGAGTG GTGATAACTT TATAATATCT 480 ATAACATTGG GTGTTACTTA TGTTGCTGTT TTTAGCATTA CACAGAGATT ATTTCAAATA 540 TCTACGGTCC CTCTTACGAT TTATAACATC CCGTTATGGG CTGCTTATGC AGATGCTCAT 600 GCACGCAATG ATACTCAATT TATAAAAAAG ACGCTCAGAA CATCATTGAA AATAGTGGGT 660 ATTTCATCAT TCTTATTGGC CTTCATATTA GTAGTGTTCG GTAGTGAAGT CGTTAATATT 720 TGGACAGAAG GAAAGATTCA GGTACCTCGA ACATTCATAA TAGCTTATGC TTTATGGTCT 780 GTTATTGATG CTTTTTCGAA TACATTTGCA AGCTTTTTAA ATGGTTTGAA CATAGTTAAA 840 CAACAAATGC TTGCTGTTGT AACATTGATA TTGATCGCAA TTCCAGCAAA ATACATCATA 900 GTTAGCCATT TTGGGTTAAC TGTTATGTTG TACTGCTTCA TTTTTATATA TATTGTAAAT 960 TACCTTATAT GGTATAAATG TAGTTTTAAA AAACATATCG ATAGACAGTT AAATATAAGA 1020 GGATGAAAAT GAAATATATA CCAGTTTACC AACCGTCATT GACAGGAAAA GAAAAAGAAT 1080 ATGTAAATGA ATGTCTGGAC TCAACGTGGA TTTCATCAAA AGGAAACTAT ATTCAGAAGT 1140 TTGAAAATAA ATTTGCGGAA CAAAACCATG TGCAATATGC AACTACTGTA AGTAATGGAA 1200 CGGTTGCTCT TCATTTAGCT TTGTTAGCGT TAGGTATATC GGAAGGAGAT GAAGTTATTG 1260 TTCCAACACT GACATATATA GCATCAGTTA ATGCTATAAA ATACACAGGA GCCACCCCCA 1320 TTTTCGTTGA TTCAGATAAT GAAACTTGGC AAATGTCTGT TAGTGACATA GAACAAAAAA 1380 TCACTAATAA AACTAAAGCT ATTATGTGTG TCCATTTATA CGGACATCCA TGTGATATGG 1440 AACAAATTGT AGAACTGGCC AAAAGTAGAA ATTTGTTTGT AATTGAAGAT TGCGCTGAAG 1500 CCTTTGGTTT TAAATATAAA GGTAAATATG TGGGAACATT TGGAGATATT TCTACTTTTA 1560 GCTTTTTTGG AAATAAAACT ATTACTACAG GTGAAGGTGG AATGGTTGTC ACGAATGACA 1620 AAACACTTTA TGACCGTTGT TTACATTTTA AAGGCCAAGG ATTAGCTGTA CATAGGCAAT 1680 ATTGGCATGA CGTTATAGGC TACAATTATA GGATGACAAA TATCTGCGCT GCTATAGGAT 1740 TAGCCCAGTT AGAACAAGCT GATGATTTTA TATCACGAAA ACGTGAAATT GCTGATATTT 1800 ATAAAAAAAA TATCAACAGT CTTGTACAAG TCCACAAGGA AAGTAAAGAT GTTTTTCACA 1860 CTTATTGGAT GGTCTCAATT CTAACTAGGA CCGCAGAGGA AAGAGAGGAA TTAAGGAATC 1920 ACCTTGCAGA TAAACTCATC GAAACAAGGC CAGTTTTTTA CCCTGTCCAC ACGATGCCAA 1980 TGTACTCGGA AAAATATCAA AAGCACCCTA TAGCTGAGGA TCTTGGTTGG CGTGGAATTA 2040 ATTTACCTAG TTTCCCCAGC CTATCGAATG AGCAAGTTAT TTATATTTGT GAATCTATTA 2100 ACGAATTTTA TAGTGATAAA TAGCCTAAAA TATTGTAAAG GTCATTCATG AAAATTGCGT 2160 TGAATTCAGA TGGATTTTAC GAGTGGGGCG GTGGAATTGA TTTTATTAAA TATATTCTGT 2220 CAATATTAGA AACGAAACCA GAAATATGTA TCGAT 2255 (2) INFORMATION FOR SEQ ID NO:2: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1026 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (A) DESCRIPTION: nucleotides 1-1026 of SEQ ID NO:l (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Escherichia coli (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2: TCGTTTGTCA TAACTGCAAT ATGCTATATT ACTTCTGGCA TGATTGATTG GCAACTAGTA 60 ATAAAAGGTA TAAACGAGAA TGTGTATGCA GAGTTACAAC ACTCAATTAA AGTCTTTGTA 120 ATCATATTTG GACTTGGAAT TTATTCAAAT GGTGTGCAAA AAGTTTATAT GGGAATACAA 180 AAAGCCTATA TAAGTAATAT TGTTAATGCC ATATTTATAT TGTTATCTAT TATTACTCTA 240 GTAATATCGT CGAAACTACA TGCGGGACTA CCAGTTTTAA TTGTCAGCAC TCTTGGTATT 300 CAATACATAT CGGGAATCTA TTTAACAATT AATCTTATTA TAAAGCGATT AATAAAGTTT 360 ACAAAAGTTA ACATACATGC TAAAAGAGAA GCTCCATATT TGATATTAAA CGGTTTTTTC 420 TTTTTTATTT TACAGTTAGG CACTCTGGCA ACATGGAGTG GTGATAACTT TATAATATCT 480 ATAACATTGG GTGTTACTTA TGTTGCTGTT TTTAGCATTA CACAGAGATT ATTTCAAATA 540 TCTACGGTCC CTCTTACGAT TTATAACATC CCGTTATGGG CTGCTTATGC AGATGCTCAT 600 GCACGCAATG ATACTCAATT TATAAAAAAG ACGCTCAGAA CATCATTGAA AATAGTGGGT 660 ATTTCATCAT TCTTATTGGC CTTCATATTA GTAGTGTTCG GTAGTGAAGT CGTTAATATT 720 TGGACAGAAG GAAAGATTCA GGTACCTCGA ACATTCATAA TAGCTTATGC TTTATGGTCT 780 GTTATTGATG CTTTTTCGAA TACATTTGCA AGCTTTTTAA ATGGTTTGAA CATAGTTAAA 840 CAACAAATGC TTGCTGTTGT AACATTGATA TTGATCGCAA TTCCAGCAAA ATACATCATA 900 GTTAGCCATT TTGGGTTAAC TGTTATGTTG TACTGCTTCA TTTTTATATA TATTGTAAAT 960 TACCTTATAT GGTATAAATG TAGTTTTAAA AAACATATCG ATAGACAGTT AAATATAAGA 1020 GGATGA 1026 (2) INFORMATION FOR SEQ ID NO:3: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1095 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (A) DESCRIPTION: nucleotides 1029-2123 of SEQ ID NO:l (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Escherichia coli (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3: ATGAAATATA TACCAGTTTA CCAACCGTCA TTGACAGGAA AAGAAAAAGA ATATGTAAAT 60 GAATGTCTGG ACTCAACGTG GATTTCATCA AAAGGAAACT ATATTCAGAA GTTTGAAAAT 120 AAATTTGCGG AACAAAACCA TGTGCAATAT GCAACTACTG TAAGTAATGG AACGGTTGCT 180 CTTCATTTAG CTTTGTTAGC GTTAGGTATA TCGGAAGGAG ATGAAGTTAT TGTTCCAACA 240 CTGACATATA TAGCATCAGT TAATGCTATA AAATACACAG GAGCCACCCC CATTTTCGTT 300 GATTCAGATA ATGAAACTTG GCAAATGTCT GTTAGTGACA TAGAACAAAA AATCACTAAT 360 AAAACTAAAG CTATTATGTG TGTCCATTTA TACGGACATC CATGTGATAT GGAACAAATT 420 GTAGAACTGG CCAAAAGTAG AAATTTGTTT GTAATTGAAG ATTGCGCTGA AGCCTTTGGT 480 TTTAAATATA AAGGTAAATA TGTGGGAACA TTTGGAGATA TTTCTACTTT TAGCTTTTTT 540 GGAAATAAAA CTATTACTAC AGGTGAAGGT GGAATGGTTG TCACGAATGA CAAAACACTT 600 TATGACCGTT GTTTACATTT TAAAGGCCAA GGATTAGCTG TACATAGGCA ATATTGGCAT 660 GACGTTATAG GCTACAATTA TAGGATGACA AATATCTGCG CTGCTATAGG ATTAGCCCAG 720 TTAGAACAAG CTGATGATTT TATATCACGA AAACGTGAAA TTGCTGATAT TTATAAAAAA 780 AATATCAACA GTCTTGTACA AGTCCACAAG GAAAGTAAAG ATGTTTTTCA CACTTATTGG 8 0 ATGGTCTCAA TTCTAACTAG GACCGCAGAG GAAAGAGAGG AATTAAGGAA TCACCTTGCA 900 GATAAACTCA TCGAAACAAG GCCAGTTTTT TACCCTGTCC ACACGATGCC AATGTACTCG 960 GAAAAATATC AAAAGCACCC TATAGCTGAG GATCTTGGTT GGCGTGGAAT TAATTTACCT 1020 AGTTTCCCCA GCCTATCGAA TGAGCAAGTT ATTTATATTT GTGAATCTAT TAACGAATTT 1080 TATAGTGATA AATAG 1095 (2) INFORMATION FOR SEQ ID NO:4: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1287 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (A) DESCRIPTION: nucleotides 913-2199 of SEQ ID NO:l (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Escherichia coli (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4: GGGTTAACTG TTATGTTGTA CTGCTTCATT TTTATATATA TTGTAAATTA CCTTATATGG 60 TATAAATGTA GTTTTAAAAA ACATATCGAT AGACAGTTAA ATATAAGAGG ATGAAAATGA 120 AATATATACC AGTTTACCAA CCGTCATTGA CAGGAAAAGA AAAAGAATAT GTAAATGAAT 180 GTCTGGACTC AACGTGGATT TCATCAAAAG GAAACTATAT TCAGAAGTTT GAAAATAAAT 240 TTGCGGAACA AAACCATGTG CAATATGCAA CTACTGTAAG TAATGGAACG GTTGCTCTTC 300 ATTTAGCTTT GTTAGCGTTA GGTATATCGG AAGGAGATGA AGTTATTGTT CCAACACTGA 360 CATATATAGC ATCAGTTAAT GCTATAAAAT ACACAGGAGC CACCCCCATT TTCGTTGATT 420 CAGATAATGA AACTTGGCAA ATGTCTGTTA.GTGACATAGA ACAAAAAATC ACTAATAAAA 480 CTAAAGCTAT TATGTGTGTC CATTTATACG GACATCCATG TGATATGGAA CAAATTGTAG 540 AACTGGCCAA AAGTAGAAAT TTGTTTGTAA TTGAAGATTG CGCTGAAGCC TTTGGTTTTA 600 AATATAAAGG TAAATATGTG GGAACATTTG GAGATATTTC TACTTTTAGC TTTTTTGGAA 660 ATAAAACTAT TACTACAGGT GAAGGTGGAA TGGTTGTCAC GAATGACAAA ACACTTTATG 720 ACCGTTGTTT ACATTTTAAA GGCCAAGGAT TAGCTGTACA TAGGCAATAT TGGCATGACG 780 TTATAGGCTA CAATTATAGG ATGACAAATA TCTGCGCTGC TATAGGATTA GCCCAGTTAG 840 AACAAGCTGA TGATTTTATA TCACGAAAAC GTGAAATTGC TGATATTTAT AAAAAAAATA 900 TCAACAGTCT TGTACAAGTC CACAAGGAAA GTAAAGATGT TTTTCACACT TATTGGATGG 960 TCTCAATTCT AACTAGGACC GCAGAGGAAA GAGAGGAATT AAGGAATCAC CTTGCAGATA 1020 AACTCATCGA AACAAGGCCA GTTTTTTACC CTGTCCACAC GATGCCAATG TACTCGGAAA 1080 AATATCAAAA GCACCCTATA GCTGAGGATC TTGGTTGGCG TGGAATTAAT TTACCTAGTT 11 0 TCCCCAGCCT ATCGAATGAG CAAGTTATTT ATATTTGTGA ATCTATTAAC GAATTTTATA 1200 GTGATAAATA GCCTAAAATA TTGTAAAGGT CATTCATGAA AATTGCGTTG AATTCAGATG 1260 GATTTTACGA GTGGGGCGGT GGAATTG 1287 (2) INFORMATION FOR SEQ ID NO:5: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 341 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (A) DESCRIPTION: amino acids encoded by nucleotides 1-1026 of SEQ ID NO:l (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Escherichia coli (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:5: Ser Phe Val He Thr Ala He Cys Tyr He Thr Ser Gly Met He Asp 1 5 10 15 Trp Gin Leu Val He Lys Gly He Asn Glu Asn Val Tyr Ala Glu Leu 20 25 30 Gin His Ser He Lys Val Phe Val He He Phe Gly Leu Gly He Tyr 35 40 45 Ser Asn Gly Val Gin Lys Val Tyr Met Gly He Gin Lys Ala Tyr He 50 55 60 Ser Asn He Val Asn Ala He Phe He Leu Leu Ser He He Thr Leu 65 70 75 80 Val He Ser Ser Lys Leu His Ala Gly Leu Pro Val Leu He Val Ser 85 90 95 Thr Leu Gly He Gin Tyr He Ser Gly He Tyr Leu Thr He Asn Leu 100 105 110 He He Lys Arg Leu He Lys Phe Thr Lys Val Asn He His Ala Lys 115 120 125 Arg Glu Ala Pro Tyr Leu He Leu Asn Gly Phe Phe Phe Phe He Leu 130 135 140 Gin Leu Gly Thr Leu Ala Thr Trp Ser Gly Asp Asn Phe He He Ser 145 150 155 160 He Thr Leu Gly Val Thr Tyr Val Ala Val Phe Ser He Thr Gin Arg 165 170 175 Leu Phe Gin He Ser Thr Val Pro Leu Thr He Tyr Asn He Pro Leu 180 185 190 Trp Ala Ala Tyr Ala Asp Ala His Ala Arg Asn Asp Thr Gin Phe He 195 200 205 Lys Lys Thr Leu Arg Thr Ser Leu Lys He Val Gly He Ser Ser Phe 210 215 220 Leu Leu Ala Phe He Leu Val Val Phe Gly Ser Glu Val Val Asn He 225 230 235 240 Trp Thr Glu Gly Lys He Gin Val Pro Arg Thr Phe He He Ala Tyr 245 250 255 Ala Leu Trp Ser Val He Asp Ala Phe Ser Asn Thr Phe Ala Ser Phe 260 265 270 Leu Asn Gly Leu Asn He Val Lys Gin Gin Met Leu Ala Val Val Thr 275 280 285 Leu He Leu He Ala He Pro Ala Lys Tyr He He Val Ser His Phe 290 295 300 Gly Leu Thr Val Met Leu Tyr Cys Phe He Phe He Tyr He Val Asn 305 310 315 320 Tyr Phe He Trp Tyr Lys Cys Ser Phe Lys Lys His He Asp Arg Gin 325 330 335 Leu Asn He Arg Gly 340 (2) INFORMATION FOR SEQ ID NO:6: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 364 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (A) DESCRIPTION: amino acids encoded by nucleotides 1029-2123 of SEQ ID NO:l (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Escherichia coli (Xi) SEQUENCE DESCRIPTION: SEQ ID NO: 6: Met Lys Tyr He Pro Val Tyr Gin Pro Ser Leu Thr Gly Lys Glu Lys 1 5 10 15 Glu Tyr Val Asn Glu Cys Leu Asp Ser Thr Trp He Ser Ser Lys Gly 20 25 30 Asn Tyr He Gin Lys Phe Glu Asn Lys Phe Ala Glu Gin Asn His Val 35 40 45 Gin Tyr Ala Thr Thr Val Ser Asn Gly Thr Val Ala Leu His Leu Ala 50 55 60 Leu Leu Ala Leu Gly He Ser Glu Gly Asp Glu Val He Val Pro Thr 65 70 75 80 Leu Thr Tyr He Ala Ser Val Asn Ala He Lys Tyr Thr Gly Ala Thr 85 90 95 Pro He Phe Val Asp Ser Asp Asn Glu Thr Trp Gin Met Ser Val Ser 100 105 110 Asp He Glu Gin Lys He Thr Asn Lys Thr Lys Ala He Met Cys Val 115 120 125 His Leu Tyr Gly His Pro Cys Asp Met Glu Gin He Val Glu Leu Ala 130 135 140 Lys Ser Arg Asn Leu Phe Val He Glu Asp Cys Ala Glu Ala Phe Gly 145 150 155 160 Ser Lys Tyr Lys Gly Lys Tyr Val Gly Thr Phe Gly Asp He Ser Thr 165 170 175 Phe Ser Phe Phe Gly Asn Lys Thr He Thr Thr Gly Glu Gly Gly Met 180 185 190 Val Val Thr Asn Asp Lys Thr Leu Tyr Asp Arg Cys Leu His Phe Lys 195 200 205 Gly Gin Gly Leu Ala Val His Arg Gin Tyr Trp His Asp Val He Gly 210 215 220 Tyr Asn Tyr Arg Met Thr Asn He Cys Ala Ala He Gly Leu Ala Gin 225 230 235 240 Leu Glu Gin Ala Asp Asp Phe He Ser Arg Lys Arg Glu He Ala Asp 245 250 255 He Tyr Lys Lys Asn He Asn Ser Leu Val Gin Val His Lys Glu Ser 260 265 270 Lys Asp Val Phe His Thr Tyr Trp Met Val Ser He Leu Thr Arg Thr 275 280 285 Ala Glu Glu Arg Glu Glu Leu Arg Asn His Leu Ala Asp Lys Leu He 290 295 300 Glu Thr Arg Pro Val Phe Tyr Pro Val His Thr Met Pro Met Tyr Ser 305 310 315 320 Glu Lys Tyr Gin Lys His Pro He Ala Glu Asp Leu Gly Trp Arg Gly 325 330 335 He Asn Leu Pro Ser Phe Pro Ser Leu Ser Asn Glu Gin Val He Tyr 340 345 350 He Cys Glu Ser He Asn Glu Phe Tyr Ser Asp Lys 355 360 (2) INFORMATION FOR SEQ ID NO:7: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 2168 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (A) DESCRIPTION: nucleotides 32-2199 of SEQ ID NO:l (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Escherichia coli (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7: CTTCTGGCAT GATTGATTGG CAACTAGTAA TAAAAGGTAT AAACGAGAAT GTGTATGCAG 60 AGTTACAACA CTCAATTAAA GTCTTTGTAA TCATATTTGG ACTTGGAATT TATTCAAATG 120 GTGTGCAAAA AGTTTATATG GGAATACAAA AAGCCTATAT AAGTAATATT GTTAATGCCA 180 TATTTATATT GTTATCTATT ATTACTCTAG TAATATCGTC GAAACTACAT GCGGGACTAC 240 CAGTTTTAAT TGTCAGCACT CTTGGTATTC AATACATATC GGGAATCTAT TTAACAATTA 300 ATCTTATTAT AAAGCGATTA ATAAAGTTTA CAAAAGTTAA CATACATGCT AAAAGAGAAG 360 CTCCATATTT GATATTAAAC GGTTTTTTCT TTTTTATTTT ACAGTTAGGC ACTCTGGCAA 420 CATGGAGTGG TGATAACTTT ATAATATCTA TAACATTGGG TGTTACTTAT GTTGCTGTTT 480 TTAGCATTAC ACAGAGATTA TTTCAAATAT CTACGGTCCC TCTTACGATT TATAACATCC 540 CGTTATGGGC TGCTTATGCA GATGCTCATG CACGCAATGA TACTCAATTT ATAAAAAAGA 600 CGCTCAGAAC ATCATTGAAA ATAGTGGGTA TTTCATCATT CTTATTGGCC TTCATATTAG 660 TAGTGTTCGG TAGTGAAGTC GTTAATATTT GGACAGAAGG AAAGATTCAG GTACCTCGAA 720 CATTCATAAT AGCTTATGCT TTATGGTCTG TTATTGATGC TTTTTCGAAT ACATTTGCAA 780 GCTTTTTAAA TGGTTTGAAC ATAGTTAAAC AACAAATGCT TGCTGTTGTA ACATTGATAT 840 TGATCGCAAT TCCAGCAAAA TACATCATAG TTAGCCATTT TGGGTTAACT GTTATGTTGT 900 ACTGCTTCAT TTTTATATAT ATTGTAAATT ACCTTATATG GTATAAATGT AGTTTTAAAA 960 AACATATCGA TAGACAGTTA AATATAAGAG GATGAAAATG AAATATATAC CAGTTTACCA 1020 ACCGTCATTG ACAGGAAAAG AAAAAGAATA TGTAAATGAA TGTCTGGACT CAACGTGGAT 1080 TTCATCAAAA GGAAACTATA TTCAGAAGTT TGAAAATAAA TTTGCGGAAC AAAACCATGT 1140 GCAATATGCA ACTACTGTAA GTAATGGAAC GGTTGCTCTT CATTTAGCTT TGTTAGCGTT 1200 AGGTATATCG GAAGGAGATG AAGTTATTGT TCCAACACTG ACATATATAG CATCAGTTAA 1260 TGCTATAAAA TACACAGGAG CCACCCCCAT TTTCGTTGAT TCAGATAATG AAACTTGGCA 1320 AATGTCTGTT AGTGACATAG AACAAAAAAT CACTAATAAA ACTAAAGCTA TTATGTGTGT 1380 CCATTTATAC GGACATCCAT GTGATATGGA ACAAATTGTA GAACTGGCCA AAAGTAGAAA 1440 TTTGTTTGTA ATTGAAGATT GCGCTGAAGC CTTTGGTTTT AAATATAAAG GTAAATATGT 1500 GGGAACATTT GGAGATATTT CTACTTTTAG CTTTTTTGGA AATAAAACTA TTACTACAGG 1560 TGAAGGTGGA ATGGTTGTCA CGAATGACAA AACACTTTAT GACCGTTGTT TACATTTTAA 1620 AGGCCAAGGA TTAGCTGTAC ATAGGCAATA TTGGCATGAC GTTATAGGCT ACAATTATAG 1680 GATGACAAAT ATCTGCGCTG CTATAGGATT AGCCCAGTTA GAACAAGCTG ATGATTTTAT 1740 ATCACGAAAA CGTGAAATTG CTGATATTTA TAAAAAAAAT ATCAACAGTC TTGTACAAGT 1800 CCACAAGGAA AGTAAAGATG TTTTTCACAC TTATTGGATG GTCTCAATTC TAACTAGGAC 1860 CGCAGAGGAA AGAGAGGAAT TAAGGAATCA CCTTGCAGAT AAACTCATCG AAACAAGGCC 1920 AGTTTTTTAC CCTGTCCACA CGATGCCAAT GTACTCGGAA AAATATCAAA AGCACCCTAT 1980 AGCTGAGGAT CTTGGTTGGC GTGGAATTAA TTTACCTAGT TTCCCCAGCC TATCGAATGA 2040 GCAAGTTATT TATATTTGTG AATCTATTAA CGAATTTTAT AGTGATAAAT AGCCTAAAAT 2100 ATTGTAAAGG TCATTCATGA AAATTGCGTT GAATTCAGAT GGATTTTACG AGTGGGGCGG 2160 TGGAATTG 2168 (2) INFORMATION FOR SEQ ID NO:8: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 21 bases (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (A) DESCRIPTION: nucleotides 32-52 of SEQ ID NO:l (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Escherichia coli (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8: CTTCTGGCAT GATTGATTGG C (2) INFORMATION FOR SEQ ID NO:9: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 20 bases (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (A) DESCRIPTION: nucleotides 2180-2199 of SEQ ID NO:l (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Escherichia coli (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9: CGAGTGGGGC GGTGGAATTG

Claims: The embodiments of the invention in which an exclusive property or privilege is claimed are defined as follows: 1. An isolated nucleic acid molecule that consists of at least 15 contiguous nucleotides of SEQ ID NO 1 or its complement. 2. A nucleic acid probe for detecting the presence of enterohemmorrhagic E. coli O157:H7, consisting of an isolated nucleic acid molecule that hybridizes stringent conditions to SEQ ID NO:l or its complement and to the DNA of E. coli O157:H7 but not the DNA of enteropathogenic E. coli O55:H7.

Inventors: BILGE SIMA S, TARR PHILLIP I, VARY JAMES C, BILGE, SIMA, S., TARR, PHILLIP, I., VARY, JAMES, C.
Applicants: BILGE SIMA S, CHILDRENS HOSP MEDICAL CENTER, TARR PHILLIP I, UNIV WASHINGTON, VARY JAMES C, BILGE, SIMA, S., CHILDREN'S HOSPITAL AND MEDICAL CENTER, TARR, PHILLIP, I., UNIVERSITY OF WASHINGTON, VARY, JAMES, C.
Application year: 1996
Main classification: C07H19
IPC codes: C12Q1