去除大肠杆菌中ColE1衍生质粒的两步法。

Two-step method for curing Escherichia coli of ColE1-derived plasmids.

作者信息

Hove-Jensen Bjarne

机构信息

Section for Biochemical Genetics, Department of Molecular Biology, University of Copenhagen, 5 Ole Maaløes Vej, Copenhagen N, Denmark.

出版信息

J Microbiol Methods. 2008 Feb;72(2):208-13. doi: 10.1016/j.mimet.2007.11.020. Epub 2007 Dec 5.

DOI:10.1016/j.mimet.2007.11.020

PMID:18164774

Abstract

To cure Escherichia coli for plasmids derived from the ColE1 replicon advantage is taken of the fact that maintenance of this replicon requires a wild-type allele of polA, encoding DNA polymerase I. Curing is achieved by cotransduction of a mutant polA allele with metE::Tn10, fadAB::Tn10 or other transposon insertions near polA. Reciprocal transduction to Met(+) Pol(+) or to Fad(+) Pol(+) ensures reestablishment of the original genotype except for loss of the plasmid. A set of useful bacterial strains is provided.

摘要

为了消除源自ColE1复制子的质粒中的大肠杆菌，利用了这样一个事实：该复制子的维持需要编码DNA聚合酶I的polA野生型等位基因。通过将突变的polA等位基因与metE::Tn10、fadAB::Tn10或polA附近的其他转座子插入进行共转导来实现消除。向Met(+) Pol(+)或Fad(+) Pol(+)的反向转导确保了除质粒丢失外原始基因型的重新建立。提供了一组有用的细菌菌株。