耻垢分枝杆菌dnaA区域与自主复制活性。
Mycobacterium smegmatis dnaA region and autonomous replication activity.
作者信息
Rajagopalan M, Qin M H, Nash D R, Madiraju M V
机构信息
Department of Microbiology, University of Texas Health Center at Tyler 75710, USA.
出版信息
J Bacteriol. 1995 Nov;177(22):6527-35. doi: 10.1128/jb.177.22.6527-6535.1995.
Abstract
Two key elements that are thought to be required for replication initiation in eubacteria are the DnaA protein, a trans-acting factor, and the replication origin, a cis-acting element. As a first step in studying the replication initiation process in mycobacteria, we have isolated a 4-kb chromosomal DNA fragment from Mycobacterium smegmatis that contains the dnaA gene. Nucleotide sequence analysis of this region revealed homologies with the rpmH gene, which codes for the ribosomal protein L34, the dnaA gene, which codes for the replication initiator protein DnaA, and the 5' end of the dnaN gene, which codes for the beta subunit of DNA polymerase III. Further, we provide evidence that when cloned into pUC18, a plasmid that is nonreplicative in M. smegmatis, the DNA fragment containing the dnaA gene and its flanking regions rendered the former capable of autonomous replication in M. smegmatis. We suggest that the M. smegmatis chromosomal origin of replication is located within the 4-kb DNA fragment.
摘要
一般认为，真细菌复制起始需要两个关键元件，即反式作用因子DnaA蛋白和顺式作用元件复制起点。作为研究分枝杆菌复制起始过程的第一步，我们从耻垢分枝杆菌中分离出一个包含dnaA基因的4kb染色体DNA片段。对该区域的核苷酸序列分析显示，它与编码核糖体蛋白L34的rpmH基因、编码复制起始蛋白DnaA的dnaA基因以及编码DNA聚合酶IIIβ亚基的dnaN基因的5'端具有同源性。此外，我们提供的证据表明，当将含有dnaA基因及其侧翼区域的DNA片段克隆到在耻垢分枝杆菌中不能自主复制的质粒pUC18中时，该片段能使pUC18在耻垢分枝杆菌中自主复制。我们认为耻垢分枝杆菌的染色体复制起点位于这个4kb的DNA片段内。
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