用于从T7启动子高效表达克隆DNA的ColE1兼容载体。

ColE1-compatible vectors for high-level expression of cloned DNAs from the T7 promoter.

作者信息

Munson M, Predki P F, Regan L

机构信息

Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06520.

出版信息

Gene. 1994 Jun 24;144(1):59-62. doi: 10.1016/0378-1119(94)90203-8.

DOI:10.1016/0378-1119(94)90203-8

PMID:8026759

Abstract

A new family of T7-based expression plasmids with unique features is described. The plasmid origin of replication (ori), derived from P15A, is compatible with that of ColE1-derived plasmids, which facilitates the co-production of proteins from these vectors and from ColE1-derived T7 expression vectors in the same cell. The plasmids are medium-copy-number and also carry the M13 ori. Consequently, both double- and single-stranded DNA can be easily obtained. The plasmids encode KmR, thus avoiding the potential for plasmid loss associated with ApR-based systems. One of the plasmids carries the lacI gene, to allow for more stringent regulation of the production of potentially toxic proteins. When the plasmids are introduced into an Escherichia coli strain such as BL21(DE3), which contains the T7 polymerase-encoding gene under control of the lacUV5 promoter, addition of IPTG initiates the production of high levels of the recombinant protein.

摘要

描述了一类具有独特特征的基于T7的新型表达质粒。该质粒的复制起点（ori）源自P15A，与源自ColE1的质粒的复制起点兼容，这有利于在同一细胞中共同表达这些载体和源自ColE1的T7表达载体的蛋白质。这些质粒为中拷贝数，并且还携带M13 ori。因此，双链和单链DNA都可以轻松获得。这些质粒编码卡那霉素抗性（KmR），从而避免了基于氨苄青霉素抗性（ApR）系统的质粒丢失的可能性。其中一个质粒携带lacI基因，以便对潜在毒性蛋白的产生进行更严格的调控。当将这些质粒导入含有在lacUV5启动子控制下的T7聚合酶编码基因的大肠杆菌菌株（如BL21(DE3)）时，加入异丙基-β-D-硫代半乳糖苷（IPTG）会启动高水平重组蛋白的产生。