Chang A C, Cohen S N
J Bacteriol. 1978 Jun;134(3):1141-56. doi: 10.1128/jb.134.3.1141-1156.1978.
Construction and characterization of a class of multicopy plasmid cloning vehicles containing the replication system of miniplasmid P15A are described. The constructed plasmids have cleavage sites within antibiotic resistance genes for a variety of commonly employed site-specific endonucleases, permitting convenient use of the insertional inactivation procedure for the selection of clones that contain hybrid DNA molecules. Although the constructed plasmids showed DNA sequence homology with the ColE1 plasmid within the replication region, were amplifiable by chloramphenicol or spectinomycin, required DNA polymerase I for replication, and shared other replication properties with ColE1, they were nevertheless compatible with ColE1. P15A-derived plasmids were not self-transmissible and were mobilized poorly by Hfr strains; however, mobilization was complemented by the presence of a ColE1 plasmid within the same cell.
本文描述了一类含有微小质粒P15A复制系统的多拷贝质粒克隆载体的构建及特性。构建的质粒在抗生素抗性基因内具有多种常用位点特异性内切酶的切割位点，便于利用插入失活程序筛选含有杂交DNA分子的克隆。尽管构建的质粒在复制区域与ColE1质粒具有DNA序列同源性，可通过氯霉素或壮观霉素扩增，复制需要DNA聚合酶I，且与ColE1具有其他复制特性，但它们与ColE1是相容的。源自P15A的质粒不能自我传递，且被高频重组（Hfr）菌株动员的能力很差；然而，同一细胞中存在ColE1质粒可补充其动员能力。
