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. 1972 Feb;69(2):466–470. doi: 10.1073/pnas.69.2.466

Evidence for Conversion of Heteroduplex Transforming DNAs to Homoduplexes by Recipient Pneumococcal Cells

Muriel Roger 1
PMCID: PMC426482  PMID: 4400650

Abstract

Heteroduplex molecules of pneumococcal DNA, prepared by cross-annealing resolved complementary strands, have been used as donors in transformation. A number of pairs of genetic markers are situated exclusively in the trans configuration in these heteroduplexes. Insertion of two markers from trans configuration into opposite DNA strands of a recipient genome should result in their segregation after one replication cycle. As a consequence, doubly transformed progeny would not appear, or would be markedly decreased. Contrary to these expectations, transformations with the heteroduplex DNAs give as many doubly transformed progeny for unlinked marker pairs as do homoduplex DNAs. For a pair of markers that normally are weakly linked, the frequencies of cotransfer are actually greater than those observed for a mixture of two singly marked homoduplex DNAs. These results lead to the conclusion that the heteroduplex DNAs are acted upon by repair enzymes of recipient cells so that markers introduced in the trans configuration are frequently converted to the cis configuration, either before or during integration. The efficiency of this conversion suggests that the repair and integration processes may be intimately connected. It is also concluded that complete breakdown of one strand of donor heteroduplex DNA does not occur during DNA uptake by recipient cells.

Keywords: DNA strand resolution, DNA repair, bacterial transformation, genetic recombination

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Selected References

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

  1. FOX M. S., ALLEN M. K. ON THE MECHANISM OF DEOXYRIBONUCLEATE INTEGRATION IN PNEUMOCOCCAL TRANSFORMATION. Proc Natl Acad Sci U S A. 1964 Aug;52:412–419. doi: 10.1073/pnas.52.2.412. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. FOX M. S., HOTCHKISS R. D. Initiation of bacterial transformation. Nature. 1957 Jun 29;179(4574):1322–1325. doi: 10.1038/1791322a0. [DOI] [PubMed] [Google Scholar]
  3. Guerrini F., Fox M. S. Genetic heterozygosity in pneumococcal transformation. Proc Natl Acad Sci U S A. 1968 Feb;59(2):429–436. doi: 10.1073/pnas.59.2.429. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Herriott R. M. FORMATION OF HETEROZYGOTES BY ANNEALING A MIXTURE OF TRANSFORMING DNAS. Proc Natl Acad Sci U S A. 1961 Feb;47(2):146–153. doi: 10.1073/pnas.47.2.146. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Herriott R. M. Structure of the hybrid transforming unit. Genetics. 1965 Dec;52(6):1235–1246. doi: 10.1093/genetics/52.6.1235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hotchkiss R. D., Gabor M. Bacterial transformation, with special reference to recombination process. Annu Rev Genet. 1970;4:193–224. doi: 10.1146/annurev.ge.04.120170.001205. [DOI] [PubMed] [Google Scholar]
  7. KENT J. L., HOTCHKISS R. D. KINETIC ANALYSIS OF MULTIPLE, LINKED RECOMBINATIONS IN PNEUMOCOCCAL TRANSFORMATION. J Mol Biol. 1964 Aug;9:308–322. doi: 10.1016/s0022-2836(64)80209-6. [DOI] [PubMed] [Google Scholar]
  8. KENT J. L., ROGER M., HOTCHKISS R. D. ON THE ROLE OF INTEGRITY OF DNA PARTICLES IN GENETIC RECOMBINATION DURING PNEUMOCOCCAL TRANSFORMATION. Proc Natl Acad Sci U S A. 1963 Oct;50:717–725. doi: 10.1073/pnas.50.4.717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. LACKS S. Molecular fate of DNA in genetic transformation of Pneumococcus. J Mol Biol. 1962 Jul;5:119–131. doi: 10.1016/s0022-2836(62)80067-9. [DOI] [PubMed] [Google Scholar]
  10. Lacks S., Greenberg B. Deoxyribonucleases of Pneumococcus. J Biol Chem. 1967 Jul 10;242(13):3108–3120. [PubMed] [Google Scholar]
  11. OPARA-KUBINSKA Z., KUBINSKI H., SZYBALSKI W. INTERACTION BETWEEN DENATURED DNA, POLYRIBONUCLEOTIDES, AND RIBOSOMAL RNA: ATTEMPTS AT PREPARATIVE SEPARATION OF THE COMPLEMENTARY DNA STRANDS. Proc Natl Acad Sci U S A. 1964 Oct;52:923–930. doi: 10.1073/pnas.52.4.923. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Peterson J. M., Guild W. R. Fractionated strands of bacterial deoxyribonucleic acid. 3. Transformation efficiencies and rates of phenotypic expression. J Bacteriol. 1968 Dec;96(6):1991–1996. doi: 10.1128/jb.96.6.1991-1996.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Roger M. Chromatographic resolution of complementary strands of denatured Pneumococcal DNA. Proc Natl Acad Sci U S A. 1968 Jan;59(1):200–207. doi: 10.1073/pnas.59.1.200. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Spatz H. C., Trautner T. A. One way to do experiments on gene conversion? Transfection with heteroduplex SPP1 DNA. Mol Gen Genet. 1970;109(1):84–106. doi: 10.1007/BF00334048. [DOI] [PubMed] [Google Scholar]
  15. Strauss N. Transformation of Bacillus subtilis using hybrid DNA molecules constructed by annealing resolved complementary strands. Genetics. 1970 Dec;66(4):583–593. doi: 10.1093/genetics/66.4.583. [DOI] [PMC free article] [PubMed] [Google Scholar]

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