Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1980 Nov;144(2):766–771. doi: 10.1128/jb.144.2.766-771.1980

Heterogeneity of deoxyribonucleic acid molecules isolated from Mycobacterium smegmatis.

M V Norgard, T Imaeda
PMCID: PMC294727  PMID: 6933148

Abstract

Bulk chromosomal deoxyribonucleic acids (DNAs) of Mycobacterium smegmatis strains 607+ (wild type) and 607-1 (Strr) and orange-red pigmented variants (OR) were separated into two distinct bands (types 1 and 2) by cesium chloride density gradient centrifugation. Thermal denaturation analyses showed that type 1 and 2 DNA fragments of these strains possessed guanine plus cytosine contents averaging 69.2% and 60.8%, respectively. Type 1 and 2 DNAs from all strains tested were recovered in relatively equal quantities upon isolation and were found to have similar molecular weights (3.0 x 10(7)). Spectrophotometric assay of DNA reassociation showed that homology between any type 1 and 2 DNA fragments was always very low (29 to 33%), even within the same strain. Homologies among type 1 DNAs isolated from any strain were always high (92 to 98%), whereas homologies between type 2 DNA isolated from OR strains and that from their parental strain 607-1 were lower (51 to 55%). Transformation experiments revealed that methionine, leucine, folic acid, and streptomycin markers were found exclusively in type 1 DNA fragments. In addition to the two types of chromosomal DNA, plasmid DNA possessing a molecular weight of about 4 x 10(6) was found in strain 607-1.

Full text

PDF
766

Images in this article

Selected References

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

  1. Bradley S. G. Relationships among mycobacteria and nocardiae based upon deoxyribonucleic acid reassociation. J Bacteriol. 1973 Feb;113(2):645–651. doi: 10.1128/jb.113.2.645-651.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bradley S. G. Significance of nucleic acid hybridization to systematics of antinomycetes. Adv Appl Microbiol. 1975;19:59–70. doi: 10.1016/s0065-2164(08)70423-4. [DOI] [PubMed] [Google Scholar]
  3. De Ley J., Cattoir H., Reynaerts A. The quantitative measurement of DNA hybridization from renaturation rates. Eur J Biochem. 1970 Jan;12(1):133–142. doi: 10.1111/j.1432-1033.1970.tb00830.x. [DOI] [PubMed] [Google Scholar]
  4. Guerry P., LeBlanc D. J., Falkow S. General method for the isolation of plasmid deoxyribonucleic acid. J Bacteriol. 1973 Nov;116(2):1064–1066. doi: 10.1128/jb.116.2.1064-1066.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hawley R. J., Imaeda T., Mann N. Isolation and characterization of nocardia-like variants of Mycobacterium smegmatis. Can J Microbiol. 1976 Oct;22(10):1480–1491. doi: 10.1139/m76-219. [DOI] [PubMed] [Google Scholar]
  6. Helinski D. R., Clewell D. B. Circular DNA. Annu Rev Biochem. 1971;40:899–942. doi: 10.1146/annurev.bi.40.070171.004343. [DOI] [PubMed] [Google Scholar]
  7. Kapp L. N., Brown S. L., Klevecz R. R. Detecting small quantities of DNA on CsCl gradients. Biochim Biophys Acta. 1974 Aug 29;361(2):140–143. doi: 10.1016/0005-2787(74)90341-4. [DOI] [PubMed] [Google Scholar]
  8. Kline B. C., Miller J. R. Detection of nonintegrated plasmid deoxyribonucleic acid in the folded chromosome of Escherichia coli: physiochemical approach to studying the unit of segregation. J Bacteriol. 1975 Jan;121(1):165–172. doi: 10.1128/jb.121.1.165-172.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Mandel M., Igambi L., Bergendahl J., Dodson M. L., Jr, Scheltgen E. Correlation of melting temperature and cesium chloride buoyant density of bacterial deoxyribonucleic acid. J Bacteriol. 1970 Feb;101(2):333–338. doi: 10.1128/jb.101.2.333-338.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Norgard M. V., Emigholz K., Monahan J. J. Increased amplification of pBR322 plasmid deoxyribonucleic acid in Escherichia coli K-12 strains RR1 and chi1776 grown in the presence of high concentrations of nucleoside. J Bacteriol. 1979 Apr;138(1):270–272. doi: 10.1128/jb.138.1.270-272.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Norgard M. V., Imaeda T. Physiological factors involved in the transformation of Mycobacterium smegmatis. J Bacteriol. 1978 Mar;133(3):1254–1262. doi: 10.1128/jb.133.3.1254-1262.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Wayne L. G., Gross W. M. Base composition of deoxyribonucleic acid isolated from mycobacteria. J Bacteriol. 1968 Dec;96(6):1915–1919. doi: 10.1128/jb.96.6.1915-1919.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Womble D. D., Taylor D. P., Rownd R. H. Method for obtaining more-accurate covalently closed circular plasmid-to-chromosome ratios from bacterial lysates by dye-buoyant density centrifugation. J Bacteriol. 1977 Apr;130(1):148–153. doi: 10.1128/jb.130.1.148-153.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES