Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1977 Jan;129(1):492–500. doi: 10.1128/jb.129.1.492-500.1977

Two-dimensional restriction analysis of the Bacillus subtilis genome: gene purification and ribosomal ribonucleic acid gene organization.

S S Potter, K F Bott, J E Newbold
PMCID: PMC234948  PMID: 401500

Abstract

With two-dimensional restriction enzyme analysis we have been able to cleave the Bacillus subtilis genome and resolve the resulting deoxyribonucleic acid (DNA) segments into discrete bands on agarose gels. A general procedure for gene purification has been developed by coupling multidimensional restriction analysis with a biological assay for gene detection. The organization of ribosomal ribonucleic acid (rRNA) genes was studied by hybridizing 16S and 23S rRNA probes to the two-dimensional DNA banding patterns.

Full text

PDF
492

Images in this article

Selected References

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

  1. Blumberg D. D., Malamy M. H. Evidence for the presence of nontranslated T7 late mRNA in infected F'(PIF+) episome-containing cells. J Virol. 1974 Feb;13(2):378–385. doi: 10.1128/jvi.13.2.378-385.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Carlton B. C., Whitt D. D. The isolation and genetic characterization of mutants of the tryptophan system of Bacillus subtilis. Genetics. 1969 Jul;62(3):445–460. doi: 10.1093/genetics/62.3.445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chow L. T., Davidson N. Electron microscope mapping of the distribution of ribosomal genes of the Bacillus subtilis chromosome. J Mol Biol. 1973 Apr 5;75(2):265–279. doi: 10.1016/0022-2836(73)90020-x. [DOI] [PubMed] [Google Scholar]
  5. Colli W., Smith I., Oishi M. Physical linkage between 5 s, 16 s and 23 s ribosomal RNA genes in Bacillus subtilis. J Mol Biol. 1971 Feb 28;56(1):117–127. doi: 10.1016/0022-2836(71)90088-x. [DOI] [PubMed] [Google Scholar]
  6. Edgell M. H., Hutchison C. A., 3rd, Sclair M. Specific endonuclease R fragments of bacteriophage phiX174 deoxyribonucleic acid. J Virol. 1972 Apr;9(4):574–582. doi: 10.1128/jvi.9.4.574-582.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Griffith J. D. Visualization of prokaryotic DNA in a regularly condensed chromatin-like fiber. Proc Natl Acad Sci U S A. 1976 Feb;73(2):563–567. doi: 10.1073/pnas.73.2.563. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Harris-Warrick R. M., Elkana Y., Ehrlich S. D., Lederberg J. Electrophoretic separation of Bacillus subtilis genes. Proc Natl Acad Sci U S A. 1975 Jun;72(6):2207–2211. doi: 10.1073/pnas.72.6.2207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Lee A. S., Sinsheimer R. L. A continuous electroelution method for the recovery of DNA restriction enzyme fragments. Anal Biochem. 1974 Aug;60(2):640–644. doi: 10.1016/0003-2697(74)90279-6. [DOI] [PubMed] [Google Scholar]
  10. Littauer U. Z., Inouye H. Regulation of tRNA. Annu Rev Biochem. 1973;42:439–470. doi: 10.1146/annurev.bi.42.070173.002255. [DOI] [PubMed] [Google Scholar]
  11. Lodish H. F. Specificity in bacterial protein synthesis: role of initiation factors and ribosomal subunits. Nature. 1970 May 23;226(5247):705–707. doi: 10.1038/226705a0. [DOI] [PubMed] [Google Scholar]
  12. Losick R., Sonenshein A. L. Change in the template specificity of RNA polymerase during sporulation of Bacillus subtilis. Nature. 1969 Oct 4;224(5214):35–37. doi: 10.1038/224035a0. [DOI] [PubMed] [Google Scholar]
  13. Lovett P. S., Bramucci M. G. Plasmid deoxyribonucleic acid in Bacillus subtilis and Bacillus pumilus. J Bacteriol. 1975 Oct;124(1):484–490. doi: 10.1128/jb.124.1.484-490.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Pettijohn D. E., Hecht R. RNA molecules bound to the folded bacterial genome stabilize DNA folds and segregate domains of supercoiling. Cold Spring Harb Symp Quant Biol. 1974;38:31–41. doi: 10.1101/sqb.1974.038.01.006. [DOI] [PubMed] [Google Scholar]
  15. Potter S. S., Newbold J. E. Multidimensional restriction enzyme analysis of complex genomes. Anal Biochem. 1976 Apr;71(2):452–458. doi: 10.1016/s0003-2697(76)80011-5. [DOI] [PubMed] [Google Scholar]
  16. Rutter W. J., Pictet R. L., Morris P. W. Toward molecular mechanisms of developmental processes. Annu Rev Biochem. 1973;42:601–646. doi: 10.1146/annurev.bi.42.070173.003125. [DOI] [PubMed] [Google Scholar]
  17. Shine J., Dalgarno L. Determinant of cistron specificity in bacterial ribosomes. Nature. 1975 Mar 6;254(5495):34–38. doi: 10.1038/254034a0. [DOI] [PubMed] [Google Scholar]
  18. Smith I., Dubnau D., Morrell P., Marmur J. Chromosomal location of DNA base sequences complementary to transfer RNA and to 5 s, 16 s and 23 s ribosomal RNA in Bacillus subtilis. J Mol Biol. 1968 Apr 14;33(1):123–140. doi: 10.1016/0022-2836(68)90285-4. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Sterlini J. M., Mandelstam J. Commitment to sporulation in Bacillus subtilis and its relationship to development of actinomycin resistance. Biochem J. 1969 Jun;113(1):29–37. doi: 10.1042/bj1130029. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Thomas M., Davis R. W. Studies on the cleavage of bacteriophage lambda DNA with EcoRI Restriction endonuclease. J Mol Biol. 1975 Jan 25;91(3):315–328. doi: 10.1016/0022-2836(75)90383-6. [DOI] [PubMed] [Google Scholar]
  22. Thuring R. W., Sanders J. P., Borst P. A freeze-squeeze method for recovering long DNA from agarose gels. Anal Biochem. 1975 May 26;66(1):213–220. doi: 10.1016/0003-2697(75)90739-3. [DOI] [PubMed] [Google Scholar]

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

RESOURCES