Isolation of a circular plasmid region sufficient for autonomous replication and transformation of infectious Borrelia burgdorferi

被引：172

作者：

Stewart, PE ^{[1
]}

Thalken, R ^{[1
]}

Bono, JL ^{[1
]}

Rosa, P ^{[1
]}

机构：

[1] NIAID, Lab Human Bacterial Pathogenesis, Rocky Mt Labs, NIH, Hamilton, MT 59840 USA

来源：

MOLECULAR MICROBIOLOGY | 2001年 / 39卷 / 03期

关键词：

D O I：

10.1046/j.1365-2958.2001.02256.x

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Borrelia burgdorferi contains abundant circular and linear plasmids, but the mechanism of replication of these extrachromosomal elements is unknown. A S. burgdorferi 9 kb circular plasmid (cp9) was amplified in its entirety by the polymerase chain reaction and used to construct a shuttle vector that replicates in Escherichia coli and B. burgdorferi. A 3.3 kb region of cp9 containing three open reading frames was used to construct a smaller shuttle vector, designated pBSV2. This vector was stably maintained in B. burgdorferi, indicating that all elements necessary for autonomous replication are probably located on this 3.3 kb fragment. a noninfectious B. Burgdorferi strain was efficiently transformed by pBSV2. Additionally, infectious B. burgdorferi was also successfully transformed by pBSV2, indicating that infectious strains of this important human pathogen can now be genetically manipulated.

引用

页码：714 / 721

页数：8

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