摘要
将棒杆菌质粒pXZ10145或pNAT65的不同酶切片段装入大肠杆菌质粒pACYCl77中构建了pTSK系列重组质粒。转化棒状类细菌的实验结果确定了质粒pXZ10145上复制必需区的位置。质粒pXZ10145复制最小必需区定位在NaeI-NruI的1.2kb片段上,在这个片段上只有一个约940碱基的阅读框架。它编码一个质粒复制因子,以对位作用方式协助那些不能自我复制但复制起始区仍保持完整的pTSK质粒在棒状类细菌中复制。质粒pXZ10145复制起始区在一个NaeI-SalI的0.3kb片段上,位于已确定的复制因子编码框架中。
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-NruI 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.
出处
《微生物学报》
CAS
CSCD
北大核心
1996年第3期168-172,共5页
Acta Microbiologica Sinica
基金
国家自然科学基金
中国科学院院重点项目
关键词
棒状类细菌
质粒
复制功能区
Coryneform bacteria, Plasmid, Minimal replicon
