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Molecular structure of uvrC gene of Escherichia coli: identification of DNA sequences required for transcription of the uvrC gene.

机译:大肠杆菌uvrC基因的分子结构:鉴定uvrC基因转录所需的DNA序列。

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摘要

We have carried out experiments to identify the regulatory regions of the uvrC gene of Escherichia coli. A uvrC+ plasmid, pUV7, containing the intact transcriptional unit for the uvrC gene, was used to subclone either the structural gene or combinations of the structural gene and 5'-flanking sequences. The plasmids so constructed were tested for ability to restore UV-resistant phenotype to uvrC- cells as an indication of expression of the uvrC gene. The chromosomal DNA in plasmid pUV7 was probed for strong binding with E. coli RNA polymerase in an attempt to identify a restriction fragment which bears the regulatory sequences for the uvrC transcriptional unit. The results indicate that DNA sequences at least 0.9 Kb upstream from the structural gene, but not the 5'-proximal sequences, regulate expression of the uvrC gene. Analysis of protein synthesis encoded by plasmid pUV7 and its derivatives suggest that there may be another gene that lies between the promoter and the uvrC gene and codes for a 27,000-Mr protein. The relation of this gene to uvrC function is not clear.
机译:我们已经进行了实验,以鉴定大肠杆菌uvrC基因的调控区。含有uvrC基因完整转录单位的uvrC +质粒pUV7用于亚克隆结构基因或结构基因与5'侧翼序列的组合。测试如此构建的质粒的恢复对uvrC-细胞的抗UV表型的能力,作为uvrC基因表达的指示。探测质粒pUV7中的染色体DNA与大肠杆菌RNA聚合酶的强结合,以试图鉴定出具有uvrC转录单位调控序列的限制性片段。结果表明,在结构基因上游至少0.9 Kb的DNA序列调节uvrC基因的表达,但不是5'-近端序列。对质粒pUV7及其衍生物编码的蛋白质合成的分析表明,在启动子和uvrC基因之间可能存在另一个基因,编码27,000-Mr蛋白。该基因与uvrC功能的关系尚不清楚。

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