NikAB- or NikB-Dependent Intracellular Recombination between Tandemly Repeated<i>oriT</i>Sequences of Plasmid R64 in Plasmid or Single-Stranded Phage Vectors

Journal of Bacteriology - Tập 185 Số 13 - Trang 3871-3877 - 2003
Nobuhisa Furuya1, Teruya Komano1
1Department of Biology, Tokyo Metropolitan University, Minamiohsawa, Hachioji, Tokyo 192-0397, Japan

Tóm tắt

ABSTRACTThe origin of transfer (oriT) of a bacterial plasmid plays a key role in both the initiation and termination of conjugative DNA transfer. We have previously shown that a conjugation-dependent recombination between the tandem R64oriTsequences cloned into pHSG398 occurred, resulting in the deletion of the intervening sequence during DNA transfer. In this study, we tandemly cloned twooriTsequences of IncI1 plasmid R64 into pUC18. Specific recombination between the twooriTsequences in pUC18 was observed withinEscherichia colicells harboring mini-R64. This recombination was found to be independent of both therecAgene and conjugative DNA transfer. The R64 genesnikAandnikB, required for conjugal DNA processing, were essential for this recombination. Although a fully active 92-bporiTsequence was required at one site for the recombination, the 44-bporiTcore sequence was sufficient at the other site. Furthermore, when twooriTsequences were tandemly cloned into the single-stranded phage vector M13 and propagated withinE. colicells, recombination between the twooriTsequences was observed, depending on thenikBgene. These results suggest that the R64 relaxase protein NikB can execute cleavage and rejoining of single-strandedoriTDNA withinE. colicells, whereas such a reaction in double-strandedoriTDNA requires collaboration of the two relaxosome proteins, NikA and NikB.

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Journal of Bacteriology

Tập 185 Số 13

3871-3877

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