Engineered promoters for finely tuned gene expression in Chlamydia

摘要 沙眼衣原体是世界范围内一个重大的健康问题。一大笔钱 负担的存在是由于患病率及其对健康和社会经济的综合影响。 急慢性疾病。衣原体是专性的细胞内寄生虫，经历 复杂的发育周期，这种生物学在历史上阻碍了 了解发病机制。最近，外源DNA转化衣原体的能力 极大地促进了对支持这一机制的分子机制的理解 衣原体感染成功。例如，衣原体基因的异位表达具有 提供了对细胞内定位和功能的洞察。随着研究变得越来越多 然而，目前复杂的启动子系统经常被证明不足以解决 与生理相关的生物学。进展尤其受到有限的选择的阻碍，因为-- 受监管的、可诱导的启动子。我们建议通过开发试剂来解决缺陷 衣原体的基因表达更加微调。我们将利用合成生物学的进展 和合理的设计，设计出一套严格监管和可调的推进器。初步 这里给出了显示该方法有效性的数据。我们建议验证和 进一步发展这些技术。在这些研究结束时，我们将建立新的 将使整个衣原体研究界受益的方法，并促进 异位基因表达用于衣原体感染生物学研究。

ABSTRACT Chlamydia trachomatis represents a significant health concern world-wide. A substantial burden exists due to the prevalence and the combined health and socioeconomic impact of acute and chronic disease. Chlamydiae are obligate intracellular parasites that undergo a complex developmental cycle, and this biology has historically impeded rapid progress in understanding pathogenesis. Recently, the ability to transform Chlamydia with exogenous DNA has greatly facilitated advances in understanding the molecular mechanisms underpinning the success of chlamydial infection. For example, ectopic expression of chlamydial genes has provided insight into intracellular localization and function. As studies become more sophisticated, however, current promoter systems are often proving inadequate to address physiologically relevant biology. Progress is particularly hindered by limited choices in tightly- regulated, inducible promoters. We propose to address deficiencies by developing reagents for more finely tuned gene expression in Chlamydia. We will exploit advances in synthetic biology and rational design to engineer a suite of tightly regulated and tunable promoters. Preliminary data showing the efficacy of the approach are presented herein. We propose to validate and further develop these technologies. At the end of these studies, we will have established new approaches that will benefit the entire Chlamydia research community and advance the utility of ectopic gene expression to study chlamydial infection biology.