用于基因组克隆的复制控制pBAC/oriV载体。

Copy-control pBAC/oriV vectors for genomic cloning.

作者信息

Wild Jadwiga, Szybalski Waclaw

机构信息

McArdle Laboratory for Cancer Research, University of Wisconsin Medical School, Madison, USA.

出版信息

Methods Mol Biol. 2004;267:145-54. doi: 10.1385/1-59259-774-2:145.

DOI:10.1385/1-59259-774-2:145

PMID:15269422

Abstract

The use of the improved BAC system for cloning genomic DNA and library constructions is described. This system retains all the advantages of the original BACs but, in addition, permits, on command, amplification of the BAC plasmids and cloned DNA. This system consists of (1) plasmid pBAC/oriV containing an additional replication origin, oriV, and (2) a host carrying the up-mutants of the trfA replicator gene expressed from the l-arabinose-inducible Para promoter. The pBAC/oriV clones are always maintained in the single-copy state, but if more DNA is required, they could be amplified up to 100-fold, depending on the size of the cloned insert.

摘要

本文描述了用于克隆基因组DNA和构建文库的改良BAC系统的应用。该系统保留了原始BAC的所有优点，此外，还可根据指令对BAC质粒和克隆的DNA进行扩增。该系统由(1)含有额外复制起点oriV的质粒pBAC/oriV和(2)携带从L-阿拉伯糖诱导型Para启动子表达的trfA复制基因上游突变体的宿主组成。pBAC/oriV克隆始终保持单拷贝状态，但如果需要更多的DNA，根据克隆插入片段的大小，它们可以扩增至100倍。