Sharma S, Dowhan W, Moses R E
Nucleic Acids Res. 1982 Sep 11;10(17):5209-21. doi: 10.1093/nar/10.17.5209.
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.
我们开展了实验以鉴定大肠杆菌uvrC基因的调控区域。一个含有uvrC基因完整转录单元的uvrC⁺质粒pUV7,被用于亚克隆结构基因,或结构基因与5'侧翼序列的组合。对如此构建的质粒进行测试,以检测其恢复uvrC⁻细胞抗紫外线表型的能力,以此作为uvrC基因表达的指标。对质粒pUV7中的染色体DNA进行检测,以寻找与大肠杆菌RNA聚合酶的强结合位点,试图鉴定出一个携带uvrC转录单元调控序列的限制性片段。结果表明,结构基因上游至少0.9 Kb的DNA序列(而非5'近端序列)调控uvrC基因的表达。对质粒pUV7及其衍生物所编码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,它编码一种分子量为27,000的蛋白质。该基因与uvrC功能的关系尚不清楚。