Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1972 May;110(2):661–666. doi: 10.1128/jb.110.2.661-666.1972

Comparison of Deoxyribonucleic Acid Uptake and Marker Integration in Bacilli and Protoplasts of Bacillus subtilis

Irving L Miller a,1, C Dwayne Palmer a, Otto E Landman a
PMCID: PMC247462  PMID: 4623311

Abstract

trp+his− donor deoxyribonucleic acid (DNA) was added to highly competent trp−his+ recipient bacilli and to protoplasts prepared from these bacilli, and the cell-DNA complexes were incubated for 30 min. The complexes were then washed and lysed, and their DNA was analyzed on a trp−his− strain for the donor marker trp+, the resident marker his+, and for the recombinant trp+his+ combination. The extracts of the bacillary complexes contained a normal percentage of donor markers (0.1-0.2%), and the number of trp+his+ doubles (20% of all trp+ transformants) indicated that the donor DNA had become integrated into the resident genomes. The protoplast complexes contained 10 to 1,000 times fewer donor markers and almost no recombinants. This indicated that, in protoplasts, marker uptake was minimal and recombination was absent. Uptake was also measured with 3H-labeled DNA. On the average, protoplasts took up one-fiftieth as much DNA as bacilli. It was concluded that, probably, protoplasts took up no DNA at all, that there were no DNA affinity sites on the surface of the protoplasts, and that the residual marker and radioactivity uptake was due to imperfections in the experimental system. The data and conclusions differed sharply from earlier ones of Hirokawa and Ikeda despite the fact that the techniques of these authors were followed in repeat experiments.

Full text

PDF
661

Selected References

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

  1. BURTON K. A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J. 1956 Feb;62(2):315–323. doi: 10.1042/bj0620315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. FOX M. S., HOTCHKISS R. D. Fate of transforming deoxyribonucleate following fixation by transformable bacteria. Nature. 1960 Sep 17;187:1002–1006. doi: 10.1038/1871002a0. [DOI] [PubMed] [Google Scholar]
  3. Hirokawa H., Ikeda Y. Genetic Recombination of Transforming Deoxyribonucleic Acid Molecules with the Recipient Genome and Among Themselves in Protoplasts of Bacillus subtilis. J Bacteriol. 1966 Aug;92(2):455–463. doi: 10.1128/jb.92.2.455-463.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Kammen H. O., Beloff R. H., Canellakis E. S. Transformation in Bacillus subtilis. I. Role of amino acids in stabilization of transformants. Biochim Biophys Acta. 1966 Jul 20;123(1):39–55. [PubMed] [Google Scholar]
  5. LANDMAN O. E., HALLE S. ENZYMICALLY AND PHYSICALLY INDUCED INHERITANCE CHANGES IN BACILLUS SUBTILIS. J Mol Biol. 1963 Dec;7:721–738. doi: 10.1016/s0022-2836(63)80119-9. [DOI] [PubMed] [Google Scholar]
  6. NESTER E. W., LEDERBERG J. Linkage of genetic units of Bacillus subtilis in DNA transformation. Proc Natl Acad Sci U S A. 1961 Jan 15;47:52–55. doi: 10.1073/pnas.47.1.52. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ranhand J. M., Theodore T. S., Cole R. M. Protein difference between competent and noncompetent cultures of a group H Streptococcus. J Bacteriol. 1970 Oct;104(1):360–362. doi: 10.1128/jb.104.1.360-362.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. SAITO H., MIURA K. I. PREPARATION OF TRANSFORMING DEOXYRIBONUCLEIC ACID BY PHENOL TREATMENT. Biochim Biophys Acta. 1963 Aug 20;72:619–629. [PubMed] [Google Scholar]
  9. STUY J. H. Transformability of Haemophilus influenzae. J Gen Microbiol. 1962 Nov;29:537–549. doi: 10.1099/00221287-29-3-537. [DOI] [PubMed] [Google Scholar]
  10. Tichy P., Landman O. E. Transformation in quasi spheroplasts of Bacillus subtilis. J Bacteriol. 1969 Jan;97(1):42–51. doi: 10.1128/jb.97.1.42-51.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Tomasz A., Zanati E. ppearance of a protein "agglutinin" on the spheroplast membrane of pneumococci during induction of competence. J Bacteriol. 1971 Mar;105(3):1213–1215. doi: 10.1128/jb.105.3.1213-1215.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. VOLL M. J., GOODGAL S. H. Recombination during transformation in Hemophilus influenzae. Proc Natl Acad Sci U S A. 1961 Apr 15;47:505–512. doi: 10.1073/pnas.47.4.505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Wilson G. A., Bott K. F. Effects of lysozyme on competence for Bacillus subtilis transfection. Biochim Biophys Acta. 1970 Feb 18;199(2):464–475. doi: 10.1016/0005-2787(70)90089-4. [DOI] [PubMed] [Google Scholar]
  14. YOUNG F. E., SPIZIZEN J. INCORPORATION OF DEOXYRIBONUCLEIC ACID IN THE BACILLUS SUBTILIS TRANSFORMATION SYSTEM. J Bacteriol. 1963 Sep;86:392–400. doi: 10.1128/jb.86.3.392-400.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES