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| An improved protein expression system for T3SS genes regulation analysis in Xanthomonas oryzae pv. oryzae |
| XU Jin-bo*, ZHANG Cui-ping*, WUNIERBIEKE Mei-li, YANG Xiao-fei, LI Yi-lang, CHEN Xiao-bin, CHEN Gong-you, ZOU Li-fang |
| Department of Environment and Resource, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, P.R.China |
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Abstract Xanthomonas oryzea pv. oryzae (Xoo) is the causal agent of bacterial blight of rice, which is a significant threat to many of rice-growing regions. The type III secretion system (T3SS) is an essential virulence factor in Xoo. Expression of the T3SS is often induced in the host environment or in hrp-inducing medium but is repressed in nutrient-rich medium. The elucidation of molecular mechanism underlying induction of T3SS genes expression is a very important step to lift the veil on global virulence regulation network in Xoo. Thus, an efficient and reliable genetic tool system is required for detection of the T3SS proteins. In this study, we constructed a protein expression vector pH3-flag based on the backbone of pHM1, a most widely used vector in Xoo strains, especially a model strain PXO99A. This vector contains a synthesized MCS-FLAG cassette that consists of a multiple cloning site (MCS), containing a modified pUC18 polylinker, and Flag as a C-terminal tag. The cassette is flanked by transcriptional terminators to eliminate interference of external transcription enabling detection of accurate protein expression. We evaluated the potential of this expression vector as T3SS proteins detection system and demonstrated it is applicable in the study of T3SS genes expression regulation in Xoo. This improved expression system could be very effectively used as a molecular tool in understanding some virulence genes expression and regulation in Xoo and other Xanthomonas spp.
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Received: 26 September 2018
Accepted:
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| Fund: This work was supported by the National Key R&D Program of China (2017YFD0200400) and the National Natural Science Foundation of China (31772122 and 31470235). |
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Corresponding Authors:
Correspondence ZOU Li-fang,
Tel: +86-21-34206149, E-mail: zoulifang202018@sjtu.edu.cn
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| About author:
XU Jin-bo, E-mail: 1140859570@qq.com; ZHANG Cui-ping,
E-mail: zylqsjzs1121@163.com; * These authors contributed equally to this study. |
Cite this article:
XU Jin-bo, ZHANG Cui-ping, WUNIERBIEKE Mei-li, YANG Xiao-fei, LI Yi-lang, CHEN Xiao-bin, CHEN Gong-you, ZOU Li-fang.
2019.
An improved protein expression system for T3SS genes regulation analysis in Xanthomonas oryzae pv. oryzae. Journal of Integrative Agriculture, 18(6): 1189-1198.
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