NikA蛋白与质粒R64 oriT区域内17个碱基对反向重复序列的一个臂的特异性结合。
Specific binding of the NikA protein to one arm of 17-base-pair inverted repeat sequences within the oriT region of plasmid R64.
作者信息
Furuya N, Komano T
机构信息
Department of Biology, Tokyo Metropolitan University, Japan.
出版信息
J Bacteriol. 1995 Jan;177(1):46-51. doi: 10.1128/jb.177.1.46-51.1995.
Abstract
Products of the nikA and nikB genes of plasmid R64 have been shown to form a relaxation complex with R64 oriT DNA and to function together as an oriT-specific nickase. We purified the protein product of the nikA gene. The purified NikA protein bound specifically to the oriT region of R64 DNA. Gel retardation assays and DNase I footprinting analyses indicated that the NikA protein bound only to the right arm of 17-bp inverted repeat sequences; the right arm differed from the left arm by a single nucleotide. The binding site is proximal to the nick site and within the 44-bp oriT core sequence. Binding of the NikA protein induced DNA bending within the R64 oriT sequence.
摘要
质粒R64的nikA和nikB基因产物已被证明可与R64 oriT DNA形成松弛复合物，并共同作为oriT特异性切口酶发挥作用。我们纯化了nikA基因的蛋白质产物。纯化的NikA蛋白特异性结合R64 DNA的oriT区域。凝胶阻滞分析和DNase I足迹分析表明，NikA蛋白仅结合17 bp反向重复序列的右臂；右臂与左臂相差一个核苷酸。结合位点靠近切口位点且位于44 bp oriT核心序列内。NikA蛋白的结合诱导了R64 oriT序列内的DNA弯曲。
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