Chromosomal duplications and cointegrates generated by the bacteriophage lamdba Red system in Escherichia coli K-12 |
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Authors: | Anthony R Poteete Anita C Fenton Ashwini Nadkarni |
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Institution: | (1) Dept. of Molecular Genetics & Microbiology, University of Massachusetts Medical School, Worcester, MA 01655, USA |
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Abstract: | Background An Escherichia coli strain in which RecBCD has been genetically replaced by the bacteriophage λ Red system engages in efficient recombination
between its chromosome and linear double-stranded DNA species sharing sequences with the chromosome. Previous studies of this
experimental system have focused on a gene replacement-type event, in which a 3.5 kbp dsDNA consisting of the cat gene and flanking lac operon sequences recombines with the E. coli chromosome to generate a chloramphenicol-resistant Lac- recombinant. The dsDNA was delivered into the cell as part of the
chromosome of a non-replicating λ vector, from which it was released by the action of a restriction endonuclease in the infected
cell. This study characterizes the genetic requirements and outcomes of a variety of additional Red-promoted homologous recombination
events producing Lac+ recombinants. |
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