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Transposon (Tn5)-mediated suppressive integration of ColE1 derivatives into the chromosome of Escherichia coli K12 (dnaA)
Authors:C Sasakawa  M Yoshikawa
Affiliation:Institute of Medical Science, University of Tokyo, Shirogane-dai, Minato-ku, Tokyo 108, Japan.
Abstract:While integration of ColE1 had not been observed previously by ordinary suppressive integration, a dnaA (Ts) E. coli strain with Tn5 at various sites of the chromosome and ColE1 or its mini-derivative, pAO3, but not pSC101, inserted by the same transposon produced integratively suppressed strains depending on the RecA function. In contrast to Hfr strains made with a stringently controlled plasmid, they contained the plasmid not only in an integrated but in an autonomous state at an amount comparable to the strain containing the plasmid only autonomously. Introduction of a RecA-deficient mutation to the strain with an integrated ColE1 derivative through conjugation failed. This is likely to be due to lethality of such a strain without RecA-dependent excision of the integrated high copy number plasmid or to quantitative deficiency of DNA polymerase I in addition to the recA mutation.
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