CONSTRUCTION AND CHARACTERIZATION OF AMPLIFIABLE MULTICOPY DNA CLONING VEHICLES DERIVED FROM P15A CRYPTIC MINIPLASMID
CONSTRUCTION AND CHARACTERIZATION OF AMPLIFIABLE MULTICOPY DNA CLONING VEHICLES DERIVED FROM P15A CRYPTIC MINIPLASMID
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DOI：
10.1128/jb.134.3.1141-1156.1978
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发表时间：
1978-01-01
影响因子：
3.2
通讯作者：
COHEN, SN
中科院分区：
文献类型：
--
作者：
CHANG, ACY;COHEN, SN
本文介绍了一类含小质粒P15A复制系统的多拷贝质粒克隆载体的构建和鉴定。所构建的质粒在抗生素耐药性基因中具有针对各种常用的位点特异性内切酶的切割位点，允许方便地使用插入失活程序来选择包含杂交DNA分子的克隆。尽管所构建的质粒在复制区与ColE1的DNA序列同源，可被氯霉素或壮观霉素扩增，需要DNA聚合酶I进行复制，并与ColE1具有其他复制特性，但它们仍与ColE1相容。P15A来源的质粒不能自我传播，不能被[EscherichiaHFR]菌株动员，但在同一细胞内存在Cole1质粒来补充动员。
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 [Escherichia coli] Hfr strains, but mobilization was complemented by the presence of a ColE1 plasmid within the same cell.