Development of a single-plasmid-based regulatable gene expression system for Borrelia burgdorferi.
Development of a single-plasmid-based regulatable gene expression system for Borrelia burgdorferi.
复制标题
开发基于单质粒的伯氏疏螺旋体可调节基因表达系统。
DOI：
10.1128/aem.02825-08
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发表时间：
2009
影响因子：
4.4
通讯作者：
Zuckert,WolframR
中科院分区：
文献类型：
--
作者：
Whetstine,ChristineR;Slusser,JoyceG;Zuckert,WolframR
We developed a single-plasmid-based regulatable protein expression system forBorrelia burgdorferi. Expression of a target gene is driven by Post, a hybridB. burgdorferi ospA-tetOpromoter, from a recombinantB. burgdorferiplasmid constitutively expressing TetR. The system was tested using the green fluorescent protein (GFP) as a reporter. Under noninducing conditions, recombinantB. burgdorfericells were nonfluorescent, no GFP protein was detected, and residual, small amounts of transcript were detectable only by reverse transcription-PCR but not by Northern blot hybridization. Upon induction with anhydrotetracycline, increasing levels of GFP transcript, protein, and fluorescence were observed. This tight and titratable promoter system will be invaluable for the study of essential borrelial proteins. Since target protein, operator, and repressor are carried by a single plasmid, the system's application is independent of a particular strain background.
