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pPSX载体的完整序列及稳定性功能分析,该载体可在大肠杆菌K12中实现链霉菌基因的稳定克隆与表达。

Complete sequence and analysis of the stability functions of pPSX, a vector that allows stable cloning and expression of Streptomycete genes in Escherichia coli K12.

作者信息

Philip Daniel S, Sarovich Derek S, Pemberton John M

机构信息

Department of Microbiology and Parasitology, University of Queensland, Brisbane 4072, Australia.

出版信息

Plasmid. 2009 Jul;62(1):39-43. doi: 10.1016/j.plasmid.2009.03.002. Epub 2009 Mar 20.

Abstract

The broad host range, cloning and expression vector pPSX has been completely sequenced and analysed. pPSX is 14.7kb in length and contains the fusion of two continuous segments of the parental 34kb, IncW plasmid pR388. pPSX appears to have retained at least three sets of gene/s which contribute in different ways to plasmid stability. The first of these parB, is a known participant in the partitioning of low-copy number plasmids. While the adjoining gene, orf35, has high homology with kfrA, a putative plasmid nucleoid organiser that is often associated with the ParAB family of proteins. The second set of genes; orfs18, 19, 20, whose exact functions are not clear, have homology to the stability operons of both IncW and IncN plasmids. The third is the resolvase, resP, which may resolve plasmid multimers that can lead to plasmid instability. pPSX is a small, stable cloning vector good for cloning and expression of a wide range of genes, including those from streptomycetes.

摘要

广宿主范围克隆与表达载体pPSX已完成全序列测定与分析。pPSX长度为14.7kb,包含亲代34kb的IncW质粒pR388两个连续片段的融合。pPSX似乎保留了至少三组以不同方式对质粒稳定性有贡献的基因。其中第一个是parB,它是已知的低拷贝数质粒分配的参与者。而相邻基因orf35与kfrA具有高度同源性,kfrA是一种假定的质粒类核组织蛋白,常与ParAB蛋白家族相关。第二组基因;orf18、orf19、orf20,其确切功能尚不清楚,与IncW和IncN质粒的稳定性操纵子具有同源性。第三个是解离酶resP,它可能会解离可导致质粒不稳定的质粒多聚体。pPSX是一种小型稳定的克隆载体,适用于克隆和表达多种基因,包括来自链霉菌的基因。

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