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. 1987 Mar;84(5):1127–1131. doi: 10.1073/pnas.84.5.1127

Cloning, sequence, and expression of the lysostaphin gene from Staphylococcus simulans.

P A Recsei, A D Gruss, R P Novick
PMCID: PMC304379  PMID: 3547405

Abstract

A 1.5-kilobase-pair fragment of DNA that contains the lysostaphin gene from Staphylococcus simulans and its flanking sequences has been cloned and completely sequenced. The gene encodes a preproenzyme of Mr 42,000. The NH2-terminal sequence of the preproenzyme is composed of a signal peptide followed by seven tandem repeats of a 13-amino acid sequence. Conversion of prolysostaphin to the mature enzyme occurs extracellularly in cultures of S. simulans and involves removal of the NH2-terminal portion of the proenzyme that contains the tandem repeats. The high degree of homology of the repeats suggests that they have arisen by duplication of a 39-base-pair sequence of DNA. In S. simulans, the lysostaphin gene is present on a large beta-lactamase plasmid.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bak A. L., Christiansen C., Stenderup A. Bacterial genome sizes determined by DNA renaturation studies. J Gen Microbiol. 1970 Dec;64(3):377–380. doi: 10.1099/00221287-64-3-377. [DOI] [PubMed] [Google Scholar]
  2. Betley M. J., Löfdahl S., Kreiswirth B. N., Bergdoll M. S., Novick R. P. Staphylococcal enterotoxin A gene is associated with a variable genetic element. Proc Natl Acad Sci U S A. 1984 Aug;81(16):5179–5183. doi: 10.1073/pnas.81.16.5179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Birnboim H. C. A rapid alkaline extraction method for the isolation of plasmid DNA. Methods Enzymol. 1983;100:243–255. doi: 10.1016/0076-6879(83)00059-2. [DOI] [PubMed] [Google Scholar]
  4. Blake M. S., Johnston K. H., Russell-Jones G. J., Gotschlich E. C. A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on Western blots. Anal Biochem. 1984 Jan;136(1):175–179. doi: 10.1016/0003-2697(84)90320-8. [DOI] [PubMed] [Google Scholar]
  5. Cisani G., Varaldo P. E., Grazi G., Soro O. High-level potentiation of lysostaphin anti-staphylococcal activity by lysozyme. Antimicrob Agents Chemother. 1982 Apr;21(4):531–535. doi: 10.1128/aac.21.4.531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Clarke L., Carbon J. A colony bank containing synthetic Col El hybrid plasmids representative of the entire E. coli genome. Cell. 1976 Sep;9(1):91–99. doi: 10.1016/0092-8674(76)90055-6. [DOI] [PubMed] [Google Scholar]
  7. Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
  8. Ersfeld-Dressen H., Sahl H. G., Brandis H. Plasmid involvement in production of and immunity to the staphylococcin-like peptide Pep 5. J Gen Microbiol. 1984 Nov;130(11):3029–3035. doi: 10.1099/00221287-130-11-3029. [DOI] [PubMed] [Google Scholar]
  9. Hollingshead S. K., Fischetti V. A., Scott J. R. Complete nucleotide sequence of type 6 M protein of the group A Streptococcus. Repetitive structure and membrane anchor. J Biol Chem. 1986 Feb 5;261(4):1677–1686. [PubMed] [Google Scholar]
  10. Iversen O. J., Grov A. Studies on lysostaphin. Separation and characterization of three enzymes. Eur J Biochem. 1973 Oct 5;38(2):293–300. doi: 10.1111/j.1432-1033.1973.tb03061.x. [DOI] [PubMed] [Google Scholar]
  11. Kloos W. E., Schleifer K. H. Simplified scheme for routine identification of human Staphylococcus species. J Clin Microbiol. 1975 Jan;1(1):82–88. doi: 10.1128/jcm.1.1.82-88.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Konisky J. Colicins and other bacteriocins with established modes of action. Annu Rev Microbiol. 1982;36:125–144. doi: 10.1146/annurev.mi.36.100182.001013. [DOI] [PubMed] [Google Scholar]
  13. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  14. Larrimore S. A., Clark S. B., Robinson J. M., Heath H. E., Sloan G. L. Coordinate production of three exoenzymes of Staphylococcus staphylolyticus. J Gen Microbiol. 1982 Jul;128(7):1529–1535. doi: 10.1099/00221287-128-7-1529. [DOI] [PubMed] [Google Scholar]
  15. Maniatis T., Jeffrey A., Kleid D. G. Nucleotide sequence of the rightward operator of phage lambda. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1184–1188. doi: 10.1073/pnas.72.3.1184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Masterson R., von David W., Wiley B. B., Rogolsky M. Mutagenesis of extrachromosomal genetic determinants for exfoliative toxin B and bacteriocin R1 synthesis in Staphylococcus aureus after plasmid transfer by protoplast fusion. Infect Immun. 1983 Dec;42(3):973–979. doi: 10.1128/iai.42.3.973-979.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  18. Mézes P. S., Blacher R. W., Lampen J. O. Processing of Bacillus cereus 569/H beta-lactamase I in Escherichia coli and Bacillus subtilis. J Biol Chem. 1985 Jan 25;260(2):1218–1223. [PubMed] [Google Scholar]
  19. Novick R. P., Brodsky R. Studies on plasmid replication. I. Plasmid incompatibility and establishment in Staphylococcus aureus. J Mol Biol. 1972 Jul 21;68(2):285–302. doi: 10.1016/0022-2836(72)90214-8. [DOI] [PubMed] [Google Scholar]
  20. Novick R. P., Murphy E., Gryczan T. J., Baron E., Edelman I. Penicillinase plasmids of Staphylococcus aureus: restriction-deletion maps. Plasmid. 1979 Jan;2(1):109–129. doi: 10.1016/0147-619x(79)90010-6. [DOI] [PubMed] [Google Scholar]
  21. O'Reilly M., Dougan G., Foster T. J., Arbuthnott J. P. Plasmids in epidermolytic strains of Staphylococcus aureus. J Gen Microbiol. 1981 May;124(1):99–107. doi: 10.1099/00221287-124-1-99. [DOI] [PubMed] [Google Scholar]
  22. O'Sullivan M. J., Marks V. Methods for the preparation of enzyme-antibody conjugates for use in enzyme immunoassay. Methods Enzymol. 1981;73(Pt B):147–166. doi: 10.1016/0076-6879(81)73062-3. [DOI] [PubMed] [Google Scholar]
  23. Poutrel B., Caffin J. P. Lysostaphin disk test for routine presumptive identification of staphylococci. J Clin Microbiol. 1981 Jun;13(6):1023–1025. doi: 10.1128/jcm.13.6.1023-1025.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Recsei P. A., Snell E. E. Histidine decarboxylase of Lactobacillus 30a. Comparative properties of wild type and mutant proenzymes and their derived enzymes. J Biol Chem. 1982 Jun 25;257(12):7196–7202. [PubMed] [Google Scholar]
  25. Robinson J. M., Hardman J. K., Sloan G. L. Relationship between lysostaphin endopeptidase production and cell wall composition in Staphylococcus staphylolyticus. J Bacteriol. 1979 Mar;137(3):1158–1164. doi: 10.1128/jb.137.3.1158-1164.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. SCHINDLER C. A., SCHUHARDT V. T. LYSOSTAPHIN: A NEW BACTERIOLYTIC AGENT FOR THE STAPHYLOCOCCUS. Proc Natl Acad Sci U S A. 1964 Mar;51:414–421. doi: 10.1073/pnas.51.3.414. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Savige W. E., Fontana A. Modification of tryptophan to oxindolylalanine by dimethyl sulfoxide-hydrochloric acid. Methods Enzymol. 1977;47:442–453. doi: 10.1016/0076-6879(77)47044-7. [DOI] [PubMed] [Google Scholar]
  29. Seidl P. H., Schleifer K. H. The immunochemistry of peptidoglycan. Antibodies against a synthetic immunogen cross-reacting with an interpeptide bridge of peptidoglycan. Eur J Biochem. 1977 Apr 1;74(2):353–363. doi: 10.1111/j.1432-1033.1977.tb11400.x. [DOI] [PubMed] [Google Scholar]
  30. Shalita Z., Murphy E., Novick R. P. Penicillinase plasmids of Staphylococcus aureus: structural and evolutionary relationships. Plasmid. 1980 May;3(3):291–311. doi: 10.1016/0147-619x(80)90042-6. [DOI] [PubMed] [Google Scholar]
  31. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  32. Springer M. S., Zanolari B. Sensory transduction in Escherichia coli: regulation of the demethylation rate by the CheA protein. Proc Natl Acad Sci U S A. 1984 Aug;81(16):5061–5065. doi: 10.1073/pnas.81.16.5061. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Stahl M. L., Ferrari E. Replacement of the Bacillus subtilis subtilisin structural gene with an In vitro-derived deletion mutation. J Bacteriol. 1984 May;158(2):411–418. doi: 10.1128/jb.158.2.411-418.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Tarr G. E. Rapid separation of amino acid phenylthiohydantoins by isocratic high-performance liquid chromatography. Anal Biochem. 1981 Feb;111(1):27–32. doi: 10.1016/0003-2697(81)90223-2. [DOI] [PubMed] [Google Scholar]
  35. Trayer H. R., Buckley C. E., 3rd Molecular properties of lysostaphin, a bacteriolytic agent specific for Staphylococcus aureus. J Biol Chem. 1970 Sep 25;245(18):4842–4846. [PubMed] [Google Scholar]
  36. Uhlén M., Guss B., Nilsson B., Gatenbeck S., Philipson L., Lindberg M. Complete sequence of the staphylococcal gene encoding protein A. A gene evolved through multiple duplications. J Biol Chem. 1984 Feb 10;259(3):1695–1702. [PubMed] [Google Scholar]
  37. Van Knippenberg P. H., Van Kimmenade J. M., Heus H. A. Phylogeny of the conserved 3' terminal structure of the RNA of small ribosomal subunits. Nucleic Acids Res. 1984 Mar 26;12(6):2595–2604. doi: 10.1093/nar/12.6.2595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Vasantha N., Thompson L. D., Rhodes C., Banner C., Nagle J., Filpula D. Genes for alkaline protease and neutral protease from Bacillus amyloliquefaciens contain a large open reading frame between the regions coding for signal sequence and mature protein. J Bacteriol. 1984 Sep;159(3):811–819. doi: 10.1128/jb.159.3.811-819.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]
  40. Watson M. E. Compilation of published signal sequences. Nucleic Acids Res. 1984 Jul 11;12(13):5145–5164. doi: 10.1093/nar/12.13.5145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Wells J. A., Ferrari E., Henner D. J., Estell D. A., Chen E. Y. Cloning, sequencing, and secretion of Bacillus amyloliquefaciens subtilisin in Bacillus subtilis. Nucleic Acids Res. 1983 Nov 25;11(22):7911–7925. doi: 10.1093/nar/11.22.7911. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Wong S. L., Price C. W., Goldfarb D. S., Doi R. H. The subtilisin E gene of Bacillus subtilis is transcribed from a sigma 37 promoter in vivo. Proc Natl Acad Sci U S A. 1984 Feb;81(4):1184–1188. doi: 10.1073/pnas.81.4.1184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Yang M. Y., Ferrari E., Henner D. J. Cloning of the neutral protease gene of Bacillus subtilis and the use of the cloned gene to create an in vitro-derived deletion mutation. J Bacteriol. 1984 Oct;160(1):15–21. doi: 10.1128/jb.160.1.15-21.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]

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