Shen T, Na S, Xiao W, Jia P
Institute of Microbiology, Academia Sinica, Beijing.
Wei Sheng Wu Xue Bao. 1996 Jun;36(3):168-72.
A pTSK series of recombinant plasmids were constructed by cloning DNA fragments of pXZ10145 or its deleted deriviate pATN65 into plasmid vector pACYC177 of E. coli. Experiment results of Coryneform bacteria transformation with these pTSK plasmids allowed us to localize the essential region for self-replication on plasmid pXZ10145. The minimal replication region of the pXZ10145 was located on a 1.2kb Nael-Nrul DNA fragment in which only one open reading frame was found. This ORF was believed to be encoded a trans-acting replication factor. The replication origin (oriV) was locate on a 0.3kb NaeI-SalI fragment which was within the ORF region.
通过将pXZ10145或其缺失衍生物pATN65的DNA片段克隆到大肠杆菌的质粒载体pACYC177中,构建了一系列pTSK重组质粒。用这些pTSK质粒对棒状杆菌进行转化的实验结果,使我们能够在质粒pXZ10145上定位自我复制的必需区域。pXZ10145的最小复制区域位于一个1.2kb的Nael - Nrul DNA片段上,在该片段中仅发现一个开放阅读框。据信这个开放阅读框编码一种反式作用复制因子。复制起点(oriV)位于一个0.3kb的NaeI - SalI片段上,该片段在开放阅读框区域内。