Orientation dependence in homologous recombination

Kenji Yamamoto, Noriko Takahashi, Youhei Fujitani, Hiroshi Yoshikura, Ichizo Kobayashi

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Homologous recombination was investigated in Escherichia coli with two plasmids, each carrying the homologous region (two defective neo genes, one with an amino-end deletion and the other with a carboxyl-end deletion) in either direct or inverted orientation. Recombination efficiency was measured in recBC sbcA and recBC sbcA strains in three ways. First, we measured the frequency of cells carrying neo+ recombinant plasmids in stationary phase. Recombination between direct repeats was much more frequent than between inverted repeats in the recBC sbcBC strain but was equally frequent in two substrates in the recBC sbcA strain. Second, the fluctuation test was used to exclude bias by a rate difference between the recombinant and parental plasmids and led to the same conclusion. Third, direct selection for recombinant just after transformation with or without substrate double- strand breaks yielded essentially the same results. Double-strand breaks elevated recombination in both the strains and in both substrates. These results are consistant with our previous findings that the major route of recombination in recBC sbcBC strains generates only one recombinant DNA from two DNAs and in recBC sbcA strains generates two recombinant DNAs from two DNAs.

Original languageEnglish
Pages (from-to)27-36
Number of pages10
JournalGenetics
Volume143
Issue number1
Publication statusPublished - 1996 May
Externally publishedYes

Fingerprint

Homologous Recombination
Genetic Recombination
Plasmids
Recombinant DNA
Nucleic Acid Repetitive Sequences
DNA
Escherichia coli
Genes

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)

Cite this

Yamamoto, K., Takahashi, N., Fujitani, Y., Yoshikura, H., & Kobayashi, I. (1996). Orientation dependence in homologous recombination. Genetics, 143(1), 27-36.

Orientation dependence in homologous recombination. / Yamamoto, Kenji; Takahashi, Noriko; Fujitani, Youhei; Yoshikura, Hiroshi; Kobayashi, Ichizo.

In: Genetics, Vol. 143, No. 1, 05.1996, p. 27-36.

Research output: Contribution to journalArticle

Yamamoto, K, Takahashi, N, Fujitani, Y, Yoshikura, H & Kobayashi, I 1996, 'Orientation dependence in homologous recombination', Genetics, vol. 143, no. 1, pp. 27-36.
Yamamoto K, Takahashi N, Fujitani Y, Yoshikura H, Kobayashi I. Orientation dependence in homologous recombination. Genetics. 1996 May;143(1):27-36.
Yamamoto, Kenji ; Takahashi, Noriko ; Fujitani, Youhei ; Yoshikura, Hiroshi ; Kobayashi, Ichizo. / Orientation dependence in homologous recombination. In: Genetics. 1996 ; Vol. 143, No. 1. pp. 27-36.
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