通过使用基因组简化的大肠杆菌MGF-01，YdfH被鉴定为rspA的阻遏物。

YdfH identified as a repressor of rspA by the use of reduced genome Escherichia coli MGF-01.

作者信息

Sakihama Yuri, Mizoguchi Hiroshi, Oshima Taku, Ogasawara Naotake

机构信息

Frontier Laboratories, Kyowa Hakko Kirin Co., Ltd., 3-6-6 Asahimachi, Machida, Tokyo 194-8533, Japan.

出版信息

Biosci Biotechnol Biochem. 2012;76(9):1688-93. doi: 10.1271/bbb.120273. Epub 2012 Sep 7.

DOI:10.1271/bbb.120273

Abstract

We have reported the construction of 1 Mb reduced genome Escherichia coli MGF-01 by a 28-step operation. This time, transcriptome analysis of MGF-01 was performed. Although the transcriptome profiles of the exponential phase in parental strain W3110red were well-conserved in MGF-01, the rspAB operon was highly expressed. A LacZ reporter assay of a series of stepwise deletion strains prepared in the course of MGF-01 construction indicated that rspA was highly expressed after the 5th step. Further analysis indicated that Δ29, one of the deleted regions at the 5th step, relates to an increase in rspA expression, and that transcriptional regulator ydfH, in the Δ29 region, is responsible for the expression of rspA, gel shift assay indicated that YdfH bound directly to the upstream region of rspA. Based on these results, it was concluded that YdfH is a transcriptional repressor of the rspAB operon.

摘要

我们曾报道通过28步操作构建了1 Mb的基因组缩减大肠杆菌MGF-01。此次，对MGF-01进行了转录组分析。尽管亲本菌株W3110red对数期的转录组图谱在MGF-01中得到了很好的保留，但rspAB操纵子却高度表达。对在MGF-01构建过程中制备的一系列逐步缺失菌株进行的LacZ报告基因检测表明，在第5步之后rspA高度表达。进一步分析表明，第5步的缺失区域之一Δ29与rspA表达的增加有关，并且Δ29区域中的转录调节因子ydfH负责rspA的表达，凝胶迁移试验表明YdfH直接结合到rspA的上游区域。基于这些结果，得出结论：YdfH是rspAB操纵子的转录阻遏物。