Construction and Expression of CpxP Protein from Plant Pathogenic Bacteria

Zi Yang, Xiaohui Zhou, Jianjun Song, Xuexing Fang, X. Xiong
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引用次数: 1

Abstract

The CpxA/R signal transduction system is one prevalent two-component systems (TCSs) in gram-negative bacteria involved in the process of bacterial adherence, invasion, motility and cell division. CpxP is the auxiliary inhibitor of Cpx-two-component system in Escherichia Coli. However, the functions of Cpx pathway in gram-negative plant pathogenic bacteria are still unknown. Previous results showed that cpxP deleted S. typhimurium is more invasive, therefore CpxP was spectulated as an inhibitor of Cpx-signalling-cascade in plant pathogenic bacteria and inhibiting its invasion to the host. To obtain the structural information of CpxP in plant pathogenic bacteria for its pathogenic research, here, for the first time, the cpxP gene from Pectobacterium carotovorum subsp. carotovorum (PC1), a plant pathogenic bacterium, was cloned into pET15b plasmid for further expression in BL21 (DE3) strain. In this study, the vector with cpxP gene was constructed which provides a way to express and purify the CpxP protein for further structural and functional studies from plant pathogenic bacteria.
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植物病原菌CpxP蛋白的构建与表达
CpxA/R信号转导系统是革兰氏阴性菌中常见的双组分系统(TCSs),参与细菌粘附、侵袭、运动和细胞分裂过程。CpxP是大肠杆菌cpx双组分体系的辅助抑制剂。然而,Cpx通路在革兰氏阴性植物病原菌中的作用尚不清楚。以往的研究结果表明,cpxP缺失的鼠伤寒沙门氏菌侵袭性更强,因此推测cpxP可能是植物致病菌cpx信号级联的抑制剂,抑制其对宿主的侵袭。为了获得植物病原菌中CpxP的结构信息,为其病原学研究提供依据,本文首次从胡萝卜乳杆菌亚群中提取CpxP基因。将植物病原菌carotovorum (PC1)克隆到pET15b质粒中,在BL21 (DE3)菌株中进一步表达。本研究构建了cpxP基因载体,为进一步从植物致病菌中表达和纯化cpxP蛋白提供了途径。
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