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The role of oriT in tra-dependent enhanced recombination between mini-F-lac-oriT and λplac5

Published online by Cambridge University Press:  14 April 2009

Jeffrey R. Carter ,
Dipty R. Patel  and
Ronald D. Porter
Jeffrey R. Carter
Affiliation:
Department of Molecular and Cell Biology, S-101 Frear Laboratory, The Pennsylvania State University, University Park, PA 16802, USA
Dipty R. Patel
Affiliation:
Department of Molecular and Cell Biology, S-101 Frear Laboratory, The Pennsylvania State University, University Park, PA 16802, USA
Ronald D. Porter*
Affiliation:
Department of Molecular and Cell Biology, S-101 Frear Laboratory, The Pennsylvania State University, University Park, PA 16802, USA
*
† Corresponding author.

Summary

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Recombination between F42lac and λplac5 is typically 20- to 50-fold more efficient than recombination between chromosomal lac and λplac5. This enhancement of recombination is recBCD-dependent and requires the expression of genes from the tra regulon of the F factor. Also required is oriT, the origin of F factor conjugational transfer, which must be located in-cis to the cellular copy of lac. In this study we show that enhanced recombination is not supported by an oriT point mutant that reduces oriT function in conjugation. We also present evidence that the activation of oriT for recombination enhancement involves the same strand-specific nick that is required for conjugal DNA transfer. Although it is thought that the role of oriT in recombination enhancement is related to the facilitated entry of RecBCD enzyme into the DNA duplex, we were unable to detect any double-strand breakage at oriT.

Type
Research Article
Information
Genetics Research , Volume 59 , Issue 3 , June 1992 , pp. 157 - 165
Copyright
Copyright © Cambridge University Press 1992

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