Transcriptional regulation mechanism of katY by OxyR in Yersinia pestis
NI Bin1,ZHANG Yi-quan2,HUANG Xin-xiang1,YANG Rui-fu2,ZHOU Dong-sheng2
1. School of Medical Science and Laboratory Medicine, Jiangsu University, Zhenjiang 212013, China; 2. State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China
Abstract：The aim of this study is to investigate the transcriptional regulation mechanism of katY by OxyR in Yersinia pestis. The entire promoter-proximal region of the katY gene was amplified by PCR from Y. pestis strain 201. And the over-expressed His-OxyR was purified under native conditions with nickel loaded HiTrap Chelating Sepharose columns (Amersham). Then, the electrophoretic mobility shift assay (EMSA) and DNase I footprinting experiment were applied to analyze the binding site of His-OxyR to katY promoter region in vitro. Total RNAs were extracted from the wide-type (WT) strain and the oxyR null mutant (ΔoxyR). Primer extension assay was employed to detect the promoter activity (the amount of primer extension products) of katY in WT and that in ΔoxyR. And then, quantitative RT-PCR was carried out to calculate the transcriptional variation of katY between the WT and ΔoxyR. The in vitro experiments showed that His-OxyR bound to a single region from 101 bp to 48 bp upstream of katY. The primer extension assay detected only one transcriptional start site located at 26 bp upstream of katY, whose transcript was under positive control of OxyR. Our data suggest that the transcription of katY is directly activated by OxyR in Yersinia pestis.
倪斌,张义全,黄新祥,杨瑞馥,周冬生. 鼠疫菌OxyR蛋白对katY的转录调控机制[J]. 中国人兽共患病学报, 2013, 29(9): 850-853.
NI Bin,ZHANG Yi-quan,HUANG Xin-xiang,YANG Rui-fu,ZHOU Dong-sheng. Transcriptional regulation mechanism of katY by OxyR in Yersinia pestis. Chinese Journal of Zoonoses, 2013, 29(9): 850-853.
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2013, Vol. 29 
