Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1985 Dec;164(3):1283–1287. doi: 10.1128/jb.164.3.1283-1287.1985

Bacteriophage SPO2-mediated plasmid transduction in transpositional mutagenesis within the genus Bacillus.

L K Kopec, R E Yasbin, R Marrero
PMCID: PMC219327  PMID: 2999078

Abstract

A single copy of the Streptococcus faecalis transposon Tn917, located in the Bacillus subtilis chromosome, was able to transpose onto the SPO2 cos plasmid pPL1017, which codes for chloramphenicol resistance and contains the bacteriophage phi 105 immunity region. Selection for pPL1017::Tn917 chimeras was performed by SPO2-mediated plasmid transduction of transposon-borne resistance to macrolide-lincosamide-streptogramin B antibiotics (MLSr). The transposition of Tn917 onto plasmid pPL1017 occurred with a frequency of 10(-5) and was dependent on the presence of a subinhibitory dose of erythromycin. Twelve chimeras were subjected to genetic and physical analyses. Two Cams transductants harbored plasmids whose chloramphenicol acetyltransferase genes had been insertionally inactivated by Tn917. Several transpositions in the vicinity of the phi 105 immunity region were detected. However, all of the 300 MLSr, Camr transductants screened were immune to phi 105 infectious activity. One pPL1017::Tn917 chimera, pLK200, was transferred by SPO2 plasmid transduction into the Bacillus amyloliquefaciens prototrophic strain DSM7. Plasmid pLK200 was effective in the mutagenesis of the DSM7 chromosome and yielded auxotrophs at a frequency of 0.5 to 5.3%. Generation of auxotrophs was also dependent on the presence of a subinhibitory dose of erythromycin. Forty-four auxotrophs representing at least nine amino acid requirements were recovered.

Full text

PDF
1283

Images in this article

1286Image
on p.1286

Selected References

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

  1. Bott K. F., Wilson G. A. Development of competence in the Bacillus subtilis transformation system. J Bacteriol. 1967 Sep;94(3):562–570. doi: 10.1128/jb.94.3.562-570.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Boylan R. J., Mendelson N. H., Brooks D., Young F. E. Regulation of the bacterial cell wall: analysis of a mutant of Bacillus subtilis defective in biosynthesis of teichoic acid. J Bacteriol. 1972 Apr;110(1):281–290. doi: 10.1128/jb.110.1.281-290.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bugaichuk U. D., Deadman M., Errington J., Savva D. Restriction enzyme analysis of Bacillus subtilis bacteriophage phi 105 DNA. J Gen Microbiol. 1984 Aug;130(8):2165–2167. doi: 10.1099/00221287-130-8-2165. [DOI] [PubMed] [Google Scholar]
  4. Cully D. F., Garro A. J. Expression of superinfection immunity to bacteriophage phi 105 by Bacillus subtilis cells carrying a plasmic chimera of pUB110 and EcoRI fragment F of phi 105 DNA. J Virol. 1980 Jun;34(3):789–791. doi: 10.1128/jvi.34.3.789-791.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dhaese P., Hussey C., Van Montagu M. Thermo-inducible gene expression in Bacillus subtilis using transcriptional regulatory elements from temperate phage phi 105. Gene. 1984 Dec;32(1-2):181–194. doi: 10.1016/0378-1119(84)90046-5. [DOI] [PubMed] [Google Scholar]
  6. Feiss M., Siegele D. A., Rudolph C. F., Frackman S. Cosmid DNA packaging in vivo. Gene. 1982 Feb;17(2):123–130. doi: 10.1016/0378-1119(82)90064-6. [DOI] [PubMed] [Google Scholar]
  7. González J. M., Jr, Dulmage H. T., Carlton B. C. Correlation between specific plasmids and delta-endotoxin production in Bacillus thuringiensis. Plasmid. 1981 May;5(3):352–365. doi: 10.1016/0147-619x(81)90010-x. [DOI] [PubMed] [Google Scholar]
  8. Gryczan T. J., Contente S., Dubnau D. Characterization of Staphylococcus aureus plasmids introduced by transformation into Bacillus subtilis. J Bacteriol. 1978 Apr;134(1):318–329. doi: 10.1128/jb.134.1.318-329.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Horinouchi S., Weisblum B. Nucleotide sequence and functional map of pC194, a plasmid that specifies inducible chloramphenicol resistance. J Bacteriol. 1982 May;150(2):815–825. doi: 10.1128/jb.150.2.815-825.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Love P. E., Lyle M. J., Yasbin R. E. DNA-damage-inducible (din) loci are transcriptionally activated in competent Bacillus subtilis. Proc Natl Acad Sci U S A. 1985 Sep;82(18):6201–6205. doi: 10.1073/pnas.82.18.6201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lovett P. S., Keggins K. M. Bacillus subtilis as a host for molecular cloning. Methods Enzymol. 1979;68:342–357. doi: 10.1016/0076-6879(79)68025-4. [DOI] [PubMed] [Google Scholar]
  12. Marrero R., Chiafari F. A., Lovett P. S. High-frequency elimination of SP02 prophage from Bacillus subtilis by plasmid transformation. J Virol. 1981 Jul;39(1):318–320. doi: 10.1128/jvi.39.1.318-320.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Marrero R., Chiafari F. A., Lovett P. S. SP02 particles mediating transduction of a plasmid containing SP02 cohesive ends. J Bacteriol. 1981 Jul;147(1):1–8. doi: 10.1128/jb.147.1.1-8.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Marrero R., Lovett P. S. Interference of plasmid pCM194 with lysogeny of bacteriophage SP02 in Bacillus subtilis. J Bacteriol. 1982 Oct;152(1):284–290. doi: 10.1128/jb.152.1.284-290.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Marrero R., Lovett P. S. Transductional selection of cloned bacteriophage phi 105 and SP02 deoxyribonucleic acids in Bacillus subtilis. J Bacteriol. 1980 Aug;143(2):879–886. doi: 10.1128/jb.143.2.879-886.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Marrero R., Young F. E., Yasbin R. E. Characterization of interspecific plasmid transfer mediated by Bacillus subtilis temperate bacteriophage SP02. J Bacteriol. 1984 Oct;160(1):458–461. doi: 10.1128/jb.160.1.458-461.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Miwa T., Matsubara K. Identification of sequences necessary for packaging DNA into lambda phage heads. Gene. 1982 Dec;20(2):267–279. doi: 10.1016/0378-1119(82)90045-2. [DOI] [PubMed] [Google Scholar]
  18. Palefski S., Hemphill H. E., Kolenbrander P. E., Whiteley H. R. Dominance relationships in mixedly infected Bacillus subtilis. J Virol. 1972 Apr;9(4):594–601. doi: 10.1128/jvi.9.4.594-601.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Spizizen J. TRANSFORMATION OF BIOCHEMICALLY DEFICIENT STRAINS OF BACILLUS SUBTILIS BY DEOXYRIBONUCLEATE. Proc Natl Acad Sci U S A. 1958 Oct 15;44(10):1072–1078. doi: 10.1073/pnas.44.10.1072. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sugino A., Higgins N. P., Brown P. O., Peebles C. L., Cozzarelli N. R. Energy coupling in DNA gyrase and the mechanism of action of novobiocin. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4838–4842. doi: 10.1073/pnas.75.10.4838. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Sullivan M. A., Yasbin R. E., Young F. E. New shuttle vectors for Bacillus subtilis and Escherichia coli which allow rapid detection of inserted fragments. Gene. 1984 Jul-Aug;29(1-2):21–26. doi: 10.1016/0378-1119(84)90161-6. [DOI] [PubMed] [Google Scholar]
  22. Tomich P. K., An F. Y., Clewell D. B. Properties of erythromycin-inducible transposon Tn917 in Streptococcus faecalis. J Bacteriol. 1980 Mar;141(3):1366–1374. doi: 10.1128/jb.141.3.1366-1374.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Tsao S. G., Brunk C. F., Pearlman R. E. Hybridization of nucleic acids directly in agarose gels. Anal Biochem. 1983 Jun;131(2):365–372. doi: 10.1016/0003-2697(83)90185-9. [DOI] [PubMed] [Google Scholar]
  24. White F. F., Klee H. J., Nester E. W. In vivo packaging of cosmids in transposon-mediated mutagenesis. J Bacteriol. 1983 Feb;153(2):1075–1078. doi: 10.1128/jb.153.2.1075-1078.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Yasbin R. E., Fields P. I., Andersen B. J. Properties of Bacillus subtilis 168 derivatives freed of their natural prophages. Gene. 1980 Dec;12(1-2):155–159. doi: 10.1016/0378-1119(80)90026-8. [DOI] [PubMed] [Google Scholar]
  26. Youngman P. J., Perkins J. B., Losick R. Genetic transposition and insertional mutagenesis in Bacillus subtilis with Streptococcus faecalis transposon Tn917. Proc Natl Acad Sci U S A. 1983 Apr;80(8):2305–2309. doi: 10.1073/pnas.80.8.2305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Youngman P., Perkins J. B., Losick R. A novel method for the rapid cloning in Escherichia coli of Bacillus subtilis chromosomal DNA adjacent to Tn917 insertions. Mol Gen Genet. 1984;195(3):424–433. doi: 10.1007/BF00341443. [DOI] [PubMed] [Google Scholar]
  28. Youngman P., Zuber P., Perkins J. B., Sandman K., Igo M., Losick R. New ways to study developmental genes in spore-forming bacteria. Science. 1985 Apr 19;228(4697):285–291. doi: 10.1126/science.228.4697.285. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES