Zaman M M, Boles T C
Graduate Department of Biochemistry, Brandeis University, Waltham, Massachusetts 02254, USA.
J Bacteriol. 1996 Jul;178(13):3840-5. doi: 10.1128/jb.178.13.3840-3845.1996.
Previously, we demonstrated that exonuclease I-deficient strains of Escherichia coli accumulate high-molecular-weight linear plasmid concatemers when transformed with plasmids carrying the chi sequence (5'- GCTGGTGG-3') (M. M. Zaman and T. C. Boles, J. Bacteriol. 176:5093-5100, 1994). Since high-molecular weight linear DNA is believed to be the natural substrate for RecBCD-mediated recombination during conjugation (A. J. Clark and K. B. Low, p. 155-215, in K. B. Low, ed., The Recombination of Genetic Material, 1988), we analyzed the recombination frequencies of chi+ and chi0 plasmids in sbcB strains. Here, we report that chi sites stimulate plasmid recombination frequency by 16-fold in sbcB strains. Chi-stimulated plasmid recombination is dependent on RecBCD but is independent of RecF pathway genes. The distribution of recombination products suggests that high-molecular-weight linear plasmid DNA is a substrate for RecBCD-mediated recombination. Surprisingly, our data also suggest that chi+ plasmids also recombine by the RecBCD pathway in rec+ sbcB+ cells.
此前,我们证明,用携带χ序列(5'-GCTGGTGG-3')的质粒转化时,核酸外切酶I缺陷型大肠杆菌菌株会积累高分子量线性质粒多联体(M. M. 扎曼和T. C. 博尔斯,《细菌学杂志》176:5093 - 5100,1994年)。由于高分子量线性DNA被认为是接合过程中RecBCD介导的重组的天然底物(A. J. 克拉克和K. B. 洛,第155 - 215页,载于K. B. 洛编,《遗传物质的重组》,1988年),我们分析了χ+和χ0质粒在sbcB菌株中的重组频率。在此,我们报告χ位点可使sbcB菌株中的质粒重组频率提高16倍。χ刺激的质粒重组依赖于RecBCD,但不依赖于RecF途径基因。重组产物的分布表明高分子量线性质粒DNA是RecBCD介导的重组的底物。令人惊讶的是,我们的数据还表明χ+质粒在rec+ sbcB+细胞中也通过RecBCD途径进行重组。