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Focused Genetic Recombination of Bacteriophage T4 Initiated by Double-Strand Breaks
Victor Shcherbakova, Igor Granovskyb, Lidiya Pluginaa, Tamara Shcherbakovaa, Svetlana Sizovaa, Konstantin Pyatkovb, Michael Shlyapnikovb, and Olga Shubinaaa Institute of Problems of Chemical Physics, Russian Academy of Sciences, Chernogolovka, Moscow Region, 142432 Russia
b Skryabin Institute of Biochemistry and Physiology of Microorganisms, RAS Pushchino, Moscow Region, 142292 Russia
Corresponding author: Victor Shcherbakov, RAS Chernogolovka, Moscow Region, 142432 Russia., svp{at}icp.ac.ru (E-mail)
Communicating editor: G. R. SMITH
strain of bacteriophage T4 was developed. The ets1, a 66-bp DNA fragment of phage T2L containing the cleavage site for the T4 SegC site-specific endonuclease, was inserted into the proximal part of the T4 rIIB gene. Under segC+ conditions, the ets1 behaves as a recombination hotspot. Crosses of the ets1 against rII markers located to the left and to the right of ets1 gave similar results, thus demonstrating the equal and symmetrical initiation of recombination by either part of the broken chromosome. Frequency/distance relationships were studied in a series of two- and three-factor crosses with other rIIB and rIIA mutants (all segC+) separated from ets1 by 12–2100 bp. The observed relationships were readily interpretable in terms of the modified splice/patch coupling model. The advantages of this localized or focused recombination over that distributed along the chromosome, as a model for studying the recombination-replication pathway in T4 in vivo, are discussed.
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  V. P. Shcherbakov, E. A. Kudryashova, T. S. Shcherbakova, S. T. Sizova, and L. A. Plugina Double-Strand Break Repair in Bacteriophage T4: Recombination Effects of 3'-5' Exonuclease Mutations Genetics, December 1, 2006; 174(4): 1729 - 1736. [Abstract] [Full Text] [PDF]
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