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. 1981 Feb;145(2):878–883. doi: 10.1128/jb.145.2.878-883.1981

Transformation of a Bacillus subtilis L-form with bacteriophage deoxyribonucleic acid.

T B White, R J Doyle, U N Streips
PMCID: PMC217193  PMID: 6780533

Abstract

A stable L-form, sal-1, of Bacillus subtilis was transformed with deoxyribonucleic acid (DNA) from bacteriophages phi 25 and phi 29 to determine whether exogenous DNA can be introduced into this organism. The viral transformation (transfection) was successful with the use of polyethylene glycol. In the presence of the fusogen, bacteriophage phi 25 DNA initiated a single cycle of infection. When compared with transfection of competent cells of Bacillus subtilis, the appearance of viral particles was delayed and their production occurred over a longer time period. L-form cells were best able to support intracellular replication of phi 25 viral particles when in balanced growth in a rich medium. The addition of polyethylene glycol also induced infection of sal-1 with whole bacteriophage phi 25 particles which could not otherwise infect the L-form and enhanced infection by intact phi 29 particles. Primary recombination was shown to be required for polyethylene glycol-mediated phi 25 transfection, but not phi 29 transfection or for whole bacteriophage phi 25 infection mediated by polyethylene glycol. Successful transfection of sal-1 suggests that the L-form may be amenable to genetic modification with exogenous DNA.

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Selected References

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

  1. Anderson D. L., Mosharrafa E. T. Physical and biological properties of phage phi 29 deoxyribonucleic acid. J Virol. 1968 Oct;2(10):1185–1190. doi: 10.1128/jvi.2.10.1185-1190.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bibb M. J., Ward J. M., Hopwood D. A. Transformation of plasmid DNA into Streptomyces at high frequency. Nature. 1978 Jul 27;274(5669):398–400. doi: 10.1038/274398a0. [DOI] [PubMed] [Google Scholar]
  3. Bott K. F., Wilson G. A. Metabolic and nutritional factors influencing the development of competence for transfection of Bacillus subtilis. Bacteriol Rev. 1968 Dec;32(4 Pt 1):370–378. [PMC free article] [PubMed] [Google Scholar]
  4. Chang S., Cohen S. N. High frequency transformation of Bacillus subtilis protoplasts by plasmid DNA. Mol Gen Genet. 1979 Jan 5;168(1):111–115. doi: 10.1007/BF00267940. [DOI] [PubMed] [Google Scholar]
  5. Delbrück M. The Burst Size Distribution in the Growth of Bacterial Viruses (Bacteriophages). J Bacteriol. 1945 Aug;50(2):131–135. [PMC free article] [PubMed] [Google Scholar]
  6. Doyle R. J., McDannel M. L., Streips U. N., Birdsell D. C., Young F. E. Polyelectrolyte nature of bacterial teichoic acids. J Bacteriol. 1974 May;118(2):606–615. doi: 10.1128/jb.118.2.606-615.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Flavell R. A., Sabo D. L., Bandle E. F., Weissmann C. Site-directed mutagenesis: generation of an extracistronic mutation in bacteriophage Q beta RNA. J Mol Biol. 1974 Oct 25;89(2):255–272. doi: 10.1016/0022-2836(74)90517-8. [DOI] [PubMed] [Google Scholar]
  8. Frehel C., Lheritier A. M., Sanchez-Rivas C., Schaeffer P. Electron microscopic study of Bacillus subtilis protoplast fusion. J Bacteriol. 1979 Mar;137(3):1354–1361. doi: 10.1128/jb.137.3.1354-1361.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. GIERER A., SCHRAMM G. Infectivity of ribonucleic acid from tobacco mosaic virus. Nature. 1956 Apr 14;177(4511):702–703. doi: 10.1038/177702a0. [DOI] [PubMed] [Google Scholar]
  10. Gilpin R. W., Young F. E., Chatterjee A. N. Characterization of a stable L-form of Bacillus subtilis 168. J Bacteriol. 1973 Jan;113(1):486–499. doi: 10.1128/jb.113.1.486-499.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Graham J. B., Istock C. A. Genetic exchange in Bacillus subtilis in soil. Mol Gen Genet. 1978 Nov 9;166(3):287–290. doi: 10.1007/BF00267620. [DOI] [PubMed] [Google Scholar]
  12. Horowitz S., Doyle R. J., Young F. E., Streips U. N. Selective association of the chromosome with membrane in a stable L-form of Bacillus subtilis. J Bacteriol. 1979 Jun;138(3):915–922. doi: 10.1128/jb.138.3.915-922.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Jacobson E. D., Landman O. E. Interaction of protoplasts, L forms, and bacilli of Bacillus subtilis with 12 strains of bacteriophage. J Bacteriol. 1975 Oct;124(1):445–448. doi: 10.1128/jb.124.1.445-448.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. KAISER A. D., HOGNESS D. S. The transformation of Escherichia coli with deoxyribonucleic acid isolated from bacteriophage lambda-dg. J Mol Biol. 1960 Dec;2:392–415. doi: 10.1016/s0022-2836(60)80050-2. [DOI] [PubMed] [Google Scholar]
  15. Liljemark W. F., Anderson D. L. Structure of Bacillus subtilis bacteriophage phi25 and phi25 deoxyribonucleic acid. J Virol. 1970 Jul;6(1):107–113. doi: 10.1128/jvi.6.1.107-113.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Murray N. E., Murray K. Manipulation of restriction targets in phage lambda to form receptor chromosomes for DNA fragments. Nature. 1974 Oct 11;251(5475):476–481. doi: 10.1038/251476a0. [DOI] [PubMed] [Google Scholar]
  17. REILLY B. E., SPIZIZEN J. BACTERIOPHAGE DEOXYRIBONUCLEATE INFECTION OF COMPETENT BACILLUS SUBTILIS. J Bacteriol. 1965 Mar;89:782–790. doi: 10.1128/jb.89.3.782-790.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. ROMIG W. R. Infection of Bacillus subtilis with phenol-extracted bacteriophages. Virology. 1962 Apr;16:452–459. doi: 10.1016/0042-6822(62)90226-x. [DOI] [PubMed] [Google Scholar]
  19. Richards G. M. Modifications of the diphenylamine reaction giving increased sensitivity and simplicity in the estimation of DNA. Anal Biochem. 1974 Feb;57(2):369–376. doi: 10.1016/0003-2697(74)90091-8. [DOI] [PubMed] [Google Scholar]
  20. SINSHEIMER R. L., STARMAN B., NAGLER C., GUTHRIE S. The process of infection with bacteriophage phi-XI74. I. Evidence for a "replicative form". J Mol Biol. 1962 Mar;4:142–160. doi: 10.1016/s0022-2836(62)80047-3. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Streips U. N., Welker N. E. Infection of Bacillus stearothermophilus with bacteriophage deoxyribonucleic acid. J Bacteriol. 1969 Jul;99(1):344–346. doi: 10.1128/jb.99.1.344-346.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Suarez J. E., Chater K. F. Polyethylene glycol-assisted transfection of Streptomyces protoplasts. J Bacteriol. 1980 Apr;142(1):8–14. doi: 10.1128/jb.142.1.8-14.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Trautner T. A., Spatz H. C. Transfection in B. subtilis. Curr Top Microbiol Immunol. 1973;62:61–88. doi: 10.1007/978-3-642-65772-6_3. [DOI] [PubMed] [Google Scholar]
  25. Wyrick P. B., Rogers H. J. Isolation and characterization of cell wall-defective variants of Bacillus subtilis and Bacillus licheniformis. J Bacteriol. 1973 Oct;116(1):456–465. doi: 10.1128/jb.116.1.456-465.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Young F. E., Haywood P., Pollock M. Isolation of L-forms of Bacillus subtilis which grow in liquid medium. J Bacteriol. 1970 Jun;102(3):867–870. doi: 10.1128/jb.102.3.867-870.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]

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