Conditional gene expression in Chlamydia trachomatis using the tet system.
Conditional gene expression in Chlamydia trachomatis using the tet system.
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DOI：
10.1371/journal.pone.0076743
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Hefty PS
中科院分区：
文献类型：
--
作者：
Wickstrum J;Sammons LR;Restivo KN;Hefty PS
沙眼衣原体是通过一个复杂的双阶段发育周期维持的，其中包含了许多鲜为人知的过程。这反映了以前遗传工具的缺乏。最近出现的一种转化衣原体的方法，使必要的分子工具得以发展，以更好地研究这些医学上重要的细菌。对衣原体生物学和发病机制的研究至关重要，是一个严格控制诱导基因表达的系统。为了实现这一目标，产生了一个新的穿梭载体，由四环素抑制因子和酸水四环素控制基因表达。该系统对GFP表达的评估表明，在诱导和去除诱导剂后恢复转录抑制后，基因调控受到严格控制，蛋白质快速表达。此外，诱导表达可以在发育周期中相对较早地被检测到，并伴随著转化为代谢活跃的衣原体形式。在发育周期的中期，观察到均匀而强烈的GFP诱导。有趣的是，在发育后期，共享包涵体内的个体生物诱导的GFP表达变化表明代谢多样性正在影响诱导和/或表达。这些观察结果支持这种分子工具的强大潜力，使许多实验分析能够更好地了解衣原体的生物学和发病机制。
Chlamydia trachomatis is maintained through a complex bi-phasic developmental cycle that incorporates numerous processes that are poorly understood. This is reflective of the previous paucity of genetic tools available. The recent advent of a method for transforming Chlamydia has enabled the development of essential molecular tools to better study these medically important bacteria. Critical for the study of Chlamydia biology and pathogenesis, is a system for tightly controlled inducible gene expression. To accomplish this, a new shuttle vector was generated with gene expression controlled by the Tetracycline repressor and anhydryotetracycline. Evaluation of GFP expression by this system demonstrated tightly controlled gene regulation with rapid protein expression upon induction and restoration of transcription repression following inducer removal. Additionally, induction of expression could be detected relatively early during the developmental cycle and concomitant with conversion into the metabolically active form of Chlamydia. Uniform and strong GFP induction was observed during middle stages of the developmental cycle. Interestingly, variable induced GFP expression by individual organisms within shared inclusions during later stages of development suggesting metabolic diversity is affecting induction and/or expression. These observations support the strong potential of this molecular tool to enable numerous experimental analyses for a better understanding of the biology and pathogenesis of Chlamydia.
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影响因子：
3.1
作者：
Song, Lihua;Carlson, John H.;Caldwell, Harlan D.
通讯作者：
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影响因子：
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