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. 1964 Sep;4(5):335–353. doi: 10.1016/s0006-3495(64)86787-4

Autoradiographic Study of Transfer of DNA during Bacterial Conjugation

Robert K Herman, Frederick Forro Jr
PMCID: PMC1367523  PMID: 14205505

Abstract

Amounts of H3-thymine-labeled DNA that are transferred from cell to cell in Hfr × F- and F+ × F- matings have been measured autoradiographically. The transferred amounts are compared to the DNA content of the bacterial chromosome, which was measured autoradiographically on the basis of a model for the replication of DNA in E. coli (1). Partition of transferred radioactivity among the progeny of F- exconjugants was also observed by autoradiography. When F+ donors were used, it was found that the transferred label often remained intact during growth of the recipients into microcolonies. The class of colonies in which this occurred was shown to contain a uniform amount of radioactivity amounting to 0.5 to 0.9 per cent of the bacterial chromosome. Dispersal of the radioactivity into smaller units was also observed. The fraction of females receiving F from F+ donors, as measured genetically, was close to the fraction of females receiving DNA-label. The distributions of amounts of radioactivity transferred to females by Hfr males after different periods of mating were shown to fit the predictions of genetic analysis of chromosomal transfer—on the assumption that the total genetic map corresponds to the whole bacterial chromosome. The label transferred by Hfr donors was found to be variably dispersed among the progeny (4 to 5 generations) of the recipient cells.

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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 T. F. Recombination and segregation in Escherichia coli. Cold Spring Harb Symp Quant Biol. 1958;23:47–58. doi: 10.1101/sqb.1958.023.01.007. [DOI] [PubMed] [Google Scholar]
  2. CAIRNS J. The bacterial chromosome and its manner of replication as seen by autoradiography. J Mol Biol. 1963 Mar;6:208–213. doi: 10.1016/s0022-2836(63)80070-4. [DOI] [PubMed] [Google Scholar]
  3. FUERST C. R., JACOB F., WOLLMAN E. L. Détermination de liaisons génétiques, chez Escherichia coli K 12, a l'aide de radiophosphore. C R Hebd Seances Acad Sci. 1956 Dec 17;243(25):2162–2164. [PubMed] [Google Scholar]
  4. GAREN A., SKAAR P. D. Transfer of phosphorus-containing material associated with mating in Escherichia coli. Biochim Biophys Acta. 1958 Mar;27(3):457–463. doi: 10.1016/0006-3002(58)90372-x. [DOI] [PubMed] [Google Scholar]
  5. JACOB F., WOLLMAN E. L. Sur les processus de conjugaison et de recombinaison génétique chez Escherichia coli. IV. Prophages inductibles et mesure des segments génétiques transferés au cours de la conjugaison. Ann Inst Pasteur (Paris) 1958 Nov;95(5):497–519. [PubMed] [Google Scholar]
  6. LAVALLE R., JACOB F. [On the sensitivity of sexual and colicinogenic episomes of Escherichia coli K12 to the disintegration of radiophosphorus]. C R Hebd Seances Acad Sci. 1961 Mar 13;252:1678–1680. [PubMed] [Google Scholar]
  7. LEVINTHAL C., THOMAS C. A., Jr Molecular autoradiography: the beta-ray counting from single virus particles and DNA molecules in nuclear emulsions. Biochim Biophys Acta. 1957 Mar;23(3):453–465. doi: 10.1016/0006-3002(57)90363-3. [DOI] [PubMed] [Google Scholar]
  8. SILVER S. D. The transfer of material during mating in Escherichia coli. Transfer of DNA and upper limits on the transfer of RNA and protein. J Mol Biol. 1963 May;6:349–360. doi: 10.1016/s0022-2836(63)80048-0. [DOI] [PubMed] [Google Scholar]
  9. SILVER S., OZEKI H. Transfer of deoxyribonucleic acid accompanying the transmission of colicinogenic properties by cell mating. Nature. 1962 Sep 1;195:873–874. doi: 10.1038/195873a0. [DOI] [PubMed] [Google Scholar]
  10. VAN TUBERGEN R. P., SETLOW R. B. Quantitative radioautographic studies on exponentially growing cultures of Escherichia coli. The distribution of parental DNA, RNA, protein, and cell wall among progeny cells. Biophys J. 1961 Sep;1:589–625. doi: 10.1016/s0006-3495(61)86911-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. WOLLMAN E. L., JACOB F. Sur le déterminisme génétique des types sexuels chez Escherichia coli K 12. C R Hebd Seances Acad Sci. 1958 Jul 28;247(4):536–539. [PubMed] [Google Scholar]

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