Rechinskiĭ V O, Savochkina L P, Bibilashvili R Sh
Mol Biol (Mosk). 1984 Jan-Feb;18(1):130-9.
Strong early bacteriophage T7 promoters A2 and A3 and also A2 and lac UV5 promoters with altered segments downstream the initiation of RNA start point were cloned using specially constructed plasmid vectors pBRS188 and PBRS240. The relative signal strengths of these promoters in vivo and in vitro were evaluated and the kinetic parameters of their interaction with RNA polymerase were determined. It has been shown that the nucleotide sequence of the transcribed region plays a significant role in specific promoter-RNA polymerase interaction and that the rate-limiting step of RNA synthesis initiation is different for various promoters.
利用特制的质粒载体pBRS188和PBRS240,克隆了强早期噬菌体T7启动子A2和A3,以及起始RNA起始点下游片段发生改变的A2和lac UV5启动子。评估了这些启动子在体内和体外的相对信号强度,并测定了它们与RNA聚合酶相互作用的动力学参数。结果表明,转录区域的核苷酸序列在特定的启动子-RNA聚合酶相互作用中起重要作用,并且不同启动子的RNA合成起始限速步骤不同。