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. 1974 Mar;13(3):584–589. doi: 10.1128/jvi.13.3.584-589.1974

Protease-Sensitive Transfection of Bacillus subtilis with Bacteriophage GA-1 DNA: a Probable Case of Heterologous Transfection

Fré Arwert 1, Gerard Venema 1
PMCID: PMC355341  PMID: 4207247

Abstract

The host bacterium of bacteriophage GA-1, Bacillus sp. G1R, was compared with respect to its taxonomic relationship to Bacillus subtilis, B. licheniformis, and B. pumilis. The physiological-biochemical properties of Bacillus sp. G1R are equal to those of B. licheniformis, but the thermal denaturation midpoint of G1R DNA differs by 3 C and the buoyant density by 0.005 g/cm3 from that of B. licheniformis. Transformation with G1R donor DNA was neither observed in B. licheniformis nor in B. subtilis-competent recipients. Bacteriophage GA-1 shows neither infectivity on B. licheniformis nor on B. subtilis. However, infection of competent B. subtilis cultures with phenol-extracted GA-1 DNA results in the production of infective GA-1 particles. The transfecting activity of GA-1 DNA is destroyed by treatment with proteolytic enzymes. Resistance of transfecting DNA to inactivation by trypsin develops earlier than that to inactivation by DNase. Protease-treated GA-1 DNA competes with transforming DNA to approximately the same extent as does untreated GA-1 DNA, suggesting that uptake of GA-1 DNA is not affected by protease treatment. CsCl density gradient centrifugation reveals that the density of trypsinized GA-1 DNA is 0.004 g/cm3 greater than that of untreated 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., Hickman D. D., Reilly B. E. Structure of Bacillus subtilis bacteriophage phi 29 and the length of phi 29 deoxyribonucleic acid. J Bacteriol. 1966 May;91(5):2081–2089. doi: 10.1128/jb.91.5.2081-2089.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Arwert F., Venema G. Transformation in Bacillus subtilis. Fate of newly introduced transforming DNA. Mol Gen Genet. 1973;123(2):185–198. doi: 10.1007/BF00267334. [DOI] [PubMed] [Google Scholar]
  4. BAYREUTHER K. E., ROMIG W. R. POLYOMA VIRUS: PRODUCTION IN BACILLUS SUBTILIS. Science. 1964 Nov 6;146(3645):778–779. doi: 10.1126/science.146.3645.778. [DOI] [PubMed] [Google Scholar]
  5. Birdsell D. C., Hathaway G. M., Rutberg L. Characterization of Temperate Bacillus Bacteriophage phi105. J Virol. 1969 Sep;4(3):264–270. doi: 10.1128/jvi.4.3.264-270.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Biswal N., Kleinschmidt A. K., Spatz H. C., Trautner T. A. Physical properties of the DNA of bacteriophage SP50. Mol Gen Genet. 1967;100(1):39–55. doi: 10.1007/BF00425774. [DOI] [PubMed] [Google Scholar]
  7. Bradley D. E. The isolation and morphology of some new bacteriophages specific for Bacillus and Acetobacter species. J Gen Microbiol. 1965 Nov;41(2):233–241. doi: 10.1099/00221287-41-2-233. [DOI] [PubMed] [Google Scholar]
  8. Bron S., Venema G. Ultraviolet inactivation and excision-repair in Bacillus subtilis. I. Construction and characterization of a transformable eightfold auxotrophic strain and two ultraviolet-sensitive derivatives. Mutat Res. 1972 May;15(1):1–10. doi: 10.1016/0027-5107(72)90086-3. [DOI] [PubMed] [Google Scholar]
  9. De Ley J. Reexamination of the association between melting point, buoyant density, and chemical base composition of deoxyribonucleic acid. J Bacteriol. 1970 Mar;101(3):738–754. doi: 10.1128/jb.101.3.738-754.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hirokawa H. Transfecting deoxyribonucleic acid of Bacillus bacteriophage phi 29 that is protease sensitive. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1555–1559. doi: 10.1073/pnas.69.6.1555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. MARMUR J., DOTY P. Determination of the base composition of deoxyribonucleic acid from its thermal denaturation temperature. J Mol Biol. 1962 Jul;5:109–118. doi: 10.1016/s0022-2836(62)80066-7. [DOI] [PubMed] [Google Scholar]
  12. Okamoto K., Mudd J. A., Mangan J., Huang W. M., Subbaiah T. V., Marmur J. Properties of the defective phage of Bacillus subtilis. J Mol Biol. 1968 Jun 28;34(3):413–428. doi: 10.1016/0022-2836(68)90169-1. [DOI] [PubMed] [Google Scholar]
  13. Ortin J., Viñuela E., Salas M., Vasquez C. DNA-protein complex in circular DNA from phage phi-29. Nat New Biol. 1971 Dec 29;234(52):275–277. doi: 10.1038/newbio234275a0. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Riva S., Polsinelli M., Falaschi A. A new phage of Bacillus subtilis with infectious DNA having separable strands. J Mol Biol. 1968 Jul 28;35(2):347–356. doi: 10.1016/s0022-2836(68)80029-4. [DOI] [PubMed] [Google Scholar]
  16. Romig W. R. Infectivity of Bacillus subtilis bacteriophage deoxyribonucleic acids extracted from mature particles and from lysogenic hosts. Bacteriol Rev. 1968 Dec;32(4 Pt 1):349–357. [PMC free article] [PubMed] [Google Scholar]
  17. Rutberg L., Hoch J. A., Spizizen J. Mechanism of transfection with deoxyribonucleic acid from the temperate Bacillus bacteriophage phi-105. J Virol. 1969 Jul;4(1):50–57. doi: 10.1128/jvi.4.1.50-57.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. SCHILDKRAUT C. L., MARMUR J., DOTY P. Determination of the base composition of deoxyribonucleic acid from its buoyant density in CsCl. J Mol Biol. 1962 Jun;4:430–443. doi: 10.1016/s0022-2836(62)80100-4. [DOI] [PubMed] [Google Scholar]
  19. STRAUSS N. CONFIGURATION OF TRANSFORMING DEOXYRIBONUCLEIC ACID DURING ENTRY INTO BACILLUS SUBTILIS. J Bacteriol. 1965 Feb;89:288–293. doi: 10.1128/jb.89.2.288-293.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Spatz H. C., Trautner T. A. The role of recombination in transfection of B. subtilis. Mol Gen Genet. 1971;113(2):174–190. doi: 10.1007/BF00333191. [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. TAYLOR M. J., THORNE C. B. TRANSDUCTION OF BACILLUS LICHENIFORMIS AND BACILLUS SUBTILIS BY EACH OF TWO PHAGES. J Bacteriol. 1963 Sep;86:452–461. doi: 10.1128/jb.86.3.452-461.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Thorne C. B., Stull H. B. Factors affecting transformation of Bacillus licheniformis. J Bacteriol. 1966 Mar;91(3):1012–1020. doi: 10.1128/jb.91.3.1012-1020.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Truffaut N., Revet B., Soulie M. O. Etude comparative des DNA de phages 2C, SP8*, SP82, phi e, SP01 et SP50. Eur J Biochem. 1970 Aug;15(2):391–400. doi: 10.1111/j.1432-1033.1970.tb01020.x. [DOI] [PubMed] [Google Scholar]
  25. Wais A. C., Goldberg E. B. Growth and transformation of phage T4 in Escherichia coli B-4, Salmonella, Aerobacter, Proteus, and Serratia. Virology. 1969 Oct;39(2):153–161. doi: 10.1016/0042-6822(69)90035-x. [DOI] [PubMed] [Google Scholar]
  26. Welker N. E., Campbell L. L. Unrelatedness of Bacillus amyloliquefaciens and Bacillus subtilis. J Bacteriol. 1967 Oct;94(4):1124–1130. doi: 10.1128/jb.94.4.1124-1130.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Williams G. L., Green D. M. Early extracellular events in infection of competent Bacillus subtilis by DNA of bacteriophage SP82G. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1545–1549. doi: 10.1073/pnas.69.6.1545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Yamamoto K. R., Alberts B. M., Benzinger R., Lawhorne L., Treiber G. Rapid bacteriophage sedimentation in the presence of polyethylene glycol and its application to large-scale virus purification. Virology. 1970 Mar;40(3):734–744. doi: 10.1016/0042-6822(70)90218-7. [DOI] [PubMed] [Google Scholar]
  29. Yelton D. B., Thorne C. B. Transduction in Bacillus cereus by each of two bacteriophages. J Bacteriol. 1970 May;102(2):573–579. doi: 10.1128/jb.102.2.573-579.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]

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