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(10) Patent No.: US 6593114 B1 USOO6593114B1 (12) United States Patent (10) Patent No.: US 6,593,114 B1 Kunsch et al. (45) Date of Patent: Jul. 15, 2003 (54) STAPHYLOCOCCUS AUREUS OTHER PUBLICATIONS POLYNUCLEOTIDES AND SEQUENCES Lewin, in Genes IV, Oxford University Press, p. 816, 1990.* (75) Inventors: Charles A. Kunsch, Norcross, GA Sharrocks, in “PCR Technology Current Innovations”, Grif (US); Gil H. Choi, Rockville, MD fin et al eds. CRC Press Inc, pp. 5-11, 1994.* (US); Steven Barash, Rockville, MD American Type Culture (ATCC), Catalogue of Bacteria & (US); Patrick J. Dillon, Carlsbad, CA Bacteriophages 17th Edition, pp. 202-204, 1989.* (US); Michael R. Fannon, Silver Walkenhorst et al, Microbiol Res, 150:347–361, 1995.* Spring, MD (US); Craig A. Rosen, Dorrell et al., Photochem. Photobiol, 58:831-835, 1993.* Laytonsville, MD (US) Sambrook et al Molecular Cloning, A Laboratory Manual, 2nd Ed., Cold Spring Harbor pp. 17.1-17.44, 1989.* (73) Assignee: Human Genome Sciences, Inc., Kennell et al, Proc. Nucl. Acid Res. Mol. Biol. 11:259-301, Rockville, MD (US) 1971.* (*) Notice: Subject to any disclaimer, the term of this Herzog et al., DNA and Cell Biology 12(6): 465-471, 1993.* ject y Jazin et al, Regulatory Peptides 47:247–258, 1993.* past Sh adjusted under 35 Rudinger et al., in “Peptide Hormones”, ed. Parsons J.A. a -- (b) by 0 days. University Park Press, pp. 1–6, 1976.* Burgess et al. et al., The Journal of Cell Biology (21) Appl. No.: 08/956,171 111:2129-2138, 1990.* (22) Filed: Oct. 20, 1997 Lazar et al., Molecular and Cellular Biology 8(3):1247–1252, 1988.* Related U.S. Application Data Jubling et al, Mol. Microbiol., 5(7): 1755–67, 1991.* (63) Continuation-in-part of application No. 08/781986, filed on * cited by examiner Jan. 3, 1997. (60) Provisional application No. 60/009,861, filed on Jan. 5, Primary Examiner Patricia A. Duffy 1996. (74) Attorney, Agent, or Firm-Human Genome Sciences, (51) Int. Cl." .......................... C12N 15/64; CO7H 21/04 ". (52) U.S. Cl. ................ 435/91.41; 435/91.4; 435/252.3; (57) ABSTRACT 435/254.11; 435/257.2; 435/320.1; 435/325; 536/23.7 The present invention provides polynucleotide Sequences of (58) Field of Search ........................ 536,237, 435/69.1, the genome of Staphylococcus aureus, polypeptide 435/69.7, 70.1, 71.1, 71.2, 320.1, 325 Sequences encoded by the polynucleotide Sequences, corre 252.3 254.11 2572. 914. 61.41: 6356. sponding polynucleotides and polypeptides, Vectors and s s s 11 12 22. 23 66 hosts comprising the polynucleotides, and assays and other s u- as a as a -1s uses thereof. The present invention further provides poly (56) References Cited nucleotide and polypeptide Sequence information Stored on computer readable media, and computer-based Systems and U.S. PATENT DOCUMENTS methods which facilitate its use. 5,175,101 A * 12/1992 Gotz et al. 6,019,984. A * 2/2000 MacInnes et al. 15 Claims, 2 Drawing Sheets US 6,593,114 B1 1 2 STAPHYLOCOCCUSAUREUS and it may reach below the skin, enter the lymphatic and POLYNUCLEOTIDES AND SEQUENCES blood circulation and develop into Septicaemia S. aureus, is among the most important pathogens typically found in burn CROSS REFERENCE TO RELATED wound infections. It can destroy granulation tissue and APPLICATIONS produce Severe Septicaemia. This application is a continuation-in-part of and claims Cellulitis priority under 35 U.S.C. S 120 to U.S. patent application Ser. Cellulitis, an acute infection of the Skin that expands from No. 08/781,986, filed Jan. 3, 1997 (pending), which is a a typically Superficial origin to spread below the cutaneous non-provisional of and claims benefit under 35 U.S.C. S layer, most commonly is caused by S. aureuS in conjunction 119(e) of U.S. Provisional Application Ser. No. 60/009,861 with S. pyrogenes. Cellulitis can lead to Systemic infection. filed Jan. 5, 1996. In fact, cellulitis can be one aspect of Synergistic bacterial Reference to a Sequence Listing Provided on Compact gangrene. This condition typically is caused by a mixture of Disc S. aureus and microaerophilic Streptococci. It causes necro sis and treatment is limited to excision of the necrotic tissue. This application refers to a "Sequence Listing”, which is 15 The condition often is fatal. provided as an electronic document on two identical com pact discs (CD-R), labeled “Copy 1” and “Copy 2.” These PEyelid infections compact discS each contain the electronic document, file S. aureus is the cause of Styes and of Sticky eye" in name “PB248P1 sequence listing..txt” (6,143,313 bytes in neonates, among other eye infections. Typically Such infec size, created on Jan. 24, 2002), which is hereby incorporated tions are limited to the Surface of the eye, and may occa in its entirety herein. Sionally penetrate the Surface with more Severe conse quences. FIELD OF THE INVENTION Food poisoning Some Strains of S. aureuS produce one or more of five The present invention relates to the field of molecular 25 Serologically distinct, heat and acid Stable enterotoxins that biology. In particular, it relates to, among other things, are not destroyed by digestive process of the Stomach and nucleotide Sequences of StaphylococcuS aureus, contigs, Small intestine (enterotoxins A-E). Ingestion of the toxin, in ORFs, fragments, probes, primers and related polynucle Sufficient quantities, typically results in Severe vomiting, but otides thereof, peptides and polypeptides encoded by the not diarrhoea. The effect does not require viable bacteria. Sequences, and uses of the polynucleotides and Sequences Although the toxins are known, their mechanism of action is thereof, Such as in fermentation, polypeptide production, not understood. assays and pharmaceutical development, among others. Joint infections BACKGROUND OF THE INVENTION S. aureuS infects bone joints causing diseaseS Such Osteo myelitis. The genus StaphylococcuS includes at least 20 distinct 35 Osteomyelitis species. (For a review see Novick, R. P., The Staphylococcus S. aureus is the most common causative agent of hae as a Molecular Genetic System, Chapter 1, pgs. 1-37 in matogenous osteomyelitis. The disease tends to occur in MOLECULAR BIOLOGY OF THE STAPHYLOCOCCI, children and adolescents more than adults and it is associ R. Novick, Ed., VCH Publishers, New York (1990)). Species ated with non-penetrating injuries to bones. Infection typi differ from one another by 80% or more, by hybridization 40 cally occurs in the long end of growing bone, hence its kinetics, whereas Strains within a species are at least 90% occurrence in physically immature populations. Most often, identical by the same measure. infection is localized in the vicinity of Sprouting capillary The Species StaphylococcuS aureus, a gram-positive, fac loops adjacent to epiphysial growth plates in the end of long, ultatively aerobic, clump-forming cocci, is among the most growing bones. important etiological agents of bacterial infection in 45 Skin infections humans, as discussed briefly below. S. aureuS is the most common pathogen of Such minor Human Health and S. Aureus skin infections as abscesses and boils. Such infections often StaphylococcuS aureus is a ubiquitous pathogen. (See, for are resolved by normal host response mechanisms, but they instance, Mims et al., MEDICAL MICROBIOLOGY, also can develop into Severe internal infections. Recurrent Mosby-Year Book Europe Limited, London, UK (1993)). It 50 infections of the nasal passages plague nasal carriers of S. is an etiological agent of a variety of conditions, ranging in LifeS . severity from mild to fatal. A few of the more common Surgical Wound Infections conditions caused by S. aureuS infection are burns, cellulitis, Surgical wounds often penetrate far into the body. Infec eyelid infections, food poisoning, joint infections, neonatal 55 tion of Such wound thus poses a grave risk to the patient. S. conjunctivitis, osteomyelitis, Skin infections, Surgical aureuS is the most important causative agent of infections in wound infection, Scalded skin Syndrome and toxic shock Surgical wounds. S. aureuS is unusually adept at invading syndrome, some of which are described further below. Surgical wounds, Sutured wounds can be infected by far Burns fewer S. aureus cells then are necessary to cause infection in Burn wounds generally are sterile initially. However, they 60 normal skin. Invasion of Surgical wound can lead to Severe generally compromise physical and immune barriers to S. aureuS Septicaemia. Invasion of the blood Stream by S. infection, cause loSS of fluid and electrolytes and result in aureuS can lead to Seeding and infection of internal organs, local or general physiological dysfunction. After cooling, particularly heart Valves and bone, causing Systemic contact with viable bacteria results in mixed colonization at diseases, Such as endocarditis and Osteomyelitis. the injury site. Infection may be restricted to the non-viable 65 Scalded Skin Syndrome debris on the burn Surface (“eschar”), it may progress into S. aureus is responsible for “scalded skin syndrome” (also full skin infection and invade viable tissue below the eschar called toxic epidermal necrosis, Ritter's disease and Lyell’s US 6,593,114 B1 3 4 disease). This diseases occurs in older children, typically in Molecular Genetics of Staphylococcus Aureus outbreaks caused by flowering of S. aureus Strains produce Despite its importance in, among other things, human exfoliation (also called Scalded skin Syndrome toxin). disease, relatively little is known about the genome of this Although the bacteria initially may infect only a minor organism. lesion, the toxin destroys intercellular connections, spreads Most genetic Studies of S. aureus have been carried out epidermal layerS and allows the infection to penetrate the using the strain NCTC8325, which contains prophages outer layer of the skin, producing the descquamation that psi 11, psi 12 and psi 13, and the UV-cured derivative of this typifies the diseases.
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  • Supplemental Table 7. Every Significant Association
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