Tsai M M, Fu Y H, Deonier R C
Department of Biological Sciences, University of Southern California, Los Angeles 90089-1340.
J Bacteriol. 1990 Aug;172(8):4603-9. doi: 10.1128/jb.172.8.4603-4609.1990.
F plasmid oriT DNA extending from the F kilobase coordinate 66.7 (base pair [bp] 1 on the oriT sequence map) rightward to bp 527 was analyzed for intrinsic bends (by permutation assays) and for binding of integration host factor (IHF) (by gel retardation and DNase footprinting). Intrinsic bending of the 527-bp fragment (bend center approximately at bp 240) was represented as a composite of at least two components located near bp 170 and near bp 260. IHF bound primarily to a site extending from bp 165 to 195 and with lower affinity to a site extending from bp 287 to 319. The intrinsic curvature and sequences to which IHF binds (IHF is known to bend DNA) may play a structural role in oriT function.
对从F千碱基坐标66.7(oriT序列图谱上的碱基对[bp]1)向右延伸至bp 527的F质粒oriT DNA进行了内在弯曲分析(通过置换分析)和整合宿主因子(IHF)结合分析(通过凝胶阻滞和DNase足迹法)。527 bp片段的内在弯曲(弯曲中心大约在bp 240处)表现为至少两个位于bp 170附近和bp 260附近的组分的组合。IHF主要结合到从bp 165延伸至195的位点,对从bp 287延伸至319的位点结合亲和力较低。IHF结合的内在曲率和序列(已知IHF可使DNA弯曲)可能在oriT功能中发挥结构作用。