Screening and application of inducible promoters in Ruminiclostridium papyrosolvens,Letters in Applied Microbiology

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Screening and application of inducible promoters in Ruminiclostridium papyrosolvens
Letters in Applied Microbiology ( IF 2.0 ) Pub Date : 2020-07-26 , DOI: 10.1111/lam.13352
Y Jiang ₁ , Y Fu ₁ , Z Ren ₂ , H Gou ₃ , C Xu ₁

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Ruminiclostridium papyrosolvens is a promising candidate for producing renewable green chemicals from cellulose due to its cellulolytic and ethanologenic capabilities. It is of significance to screen effective, and convenient‐to‐use inducible promoters that can be used for regulating the gene expression in R. papyrosolvens. We characterized two endogenous inducible promoters and investigated another two exogenous ones on the adaptability in R. papyrosolvens. Both of the endogenous xylan‐inducible promoter Pxyl and exogenous lactose‐inducible promoter Plac are found of high specificity and stringency. Pxyl has a short time to be induced while Plac has a low concentration of inducer. With these findings, a mazF‐based counter selectable system has been constructed for promoting the efficiency of mutant screening via plasmid curing. The inducible gene expression systems provided novel tools for enhancing the capability of genetic manipulation in engineering R. papyrosolvens.

中文翻译：

Ruminiclostridium papyrosolvens中诱导型启动子的筛选及应用

Ruminiclostridium papyrosolvens 由于其纤维素分解和乙醇生成能力，是从纤维素生产可再生绿色化学品的有希望的候选者。筛选出有效且使用方便的诱导型启动子，可用于调控 R. papyrosolvens 中的基因表达具有重要意义。我们表征了两个内源性诱导型启动子，并研究了另外两个外源性启动子对 R. papyrosolvens 的适应性。内源性木聚糖诱导型启动子 Pxyl 和外源性乳糖诱导型启动子 Plac 均具有高度特异性和严格性。Pxyl 的诱导时间较短，而 Plac 的诱导剂浓度较低。根据这些发现，构建了一个基于 mazF 的计数器选择系统，用于通过质粒固化提高突变体筛选的效率。

更新日期：2020-07-26

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