| 十字花科黑腐病菌hrpG基因原核表达 |
| |
| 引用本文： | 冯国芳,袁夏莹,陆光涛.十字花科黑腐病菌hrpG基因原核表达[J].广西农业生物科学,2011,30(5):577-582. |
| |
| 作者姓名： | 冯国芳 袁夏莹 陆光涛 |
| |
| 作者单位： | 广西大学生命科学与技术学院,南宁,530005 |
| |
| 基金项目： | 国家自然科学基金项目(30771177)资助
致谢 本研究在实验构思方面承蒙广西大学生命科学与技术学院苏辉昭博士、李瑞芳博士和张文飞博士的悉心指导;特致感谢! |
| |
| 摘 要： | 在十字花科黑腐病菌（Xcc）中,hrp基因对寄主的致病性和非寄主的超敏反应中起核心作用,而hrpG对整个hrp基因簇起调控作用。HrpG为OmpR家族的双组分系统感受调控蛋白,含有两个结构域,分别是N端Response_reg和C端Trans reg_C。本研究利用表达载体pQE-30Xa,成功构建了HrpG的表达重组子,在E.coliM15pREP4]中进行诱导表达。通过调节诱导温度、IPTG浓度和诱导时间最终确定在温度为20℃,IPTG浓度为0.8mmol/L,诱导表达4h。hrpG基因在宿主细胞E.coli M15获得高效可溶性表达。目前尚未有可溶性HrpG蛋白获得成功表达的报导,本研究中获得HrpG蛋白在大肠杆菌获得大量可溶性的表达,将为in vitro研究HrpG的生理活性,特异的结合位点和调控功能研究打下良好基础。
|
| 关 键 词： | 十字花科黑腐病菌 HrpG 结构特性 原核表达 |
Prokaryotic Expression of hrpG Gene from Xanthomonas campestris pv.campestris |
| |
| Feng Guofang,Yuan Xiaying,Lu Guangtao.Prokaryotic Expression of hrpG Gene from Xanthomonas campestris pv.campestris[J].Journal of Guangxi Agricultural and Biological Science,2011,30(5):577-582. |
| |
| Authors: | Feng Guofang Yuan Xiaying Lu Guangtao |
| |
| Institution: | 1,2＊ 1 College of Life Science and Technology,Guangxi University,Nanning,530005;2 State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources,Nanning,530005 |
| |
| Abstract: | hrp cluster is regulated by hrpG,which plays an important core role in invasion and non-host hypersensitive response to pathogesis host.HrpG is one member of the OmpR family response regulator with two conserved domains,which are the_reg at N terminal and the Trans reg_C at C terminal,respectively.Architecture characteristic analysis shows that these two domains are quite different from each other.In this study,we succeed to construct the recombinant plasmid to express the gene of hrpG in E.coli M15pREP4],when the sequence of hrpG gene was inserted into the expression vector of pQE-30Xa.Up to now,there is no report about the expression of soluble HrpG protein.In order to get the soluble HrpG protein with a higher yield,the reaction of expression of the hrpG protein was optimized by adjusting the tempreture and the concentration of IPTG and the time for induction respectively.The result show the best optimum condition for of the soluble HrpG production in supernatant of cell disrution is induced at 20℃ for 4 h with IPTG concentration 0.8 mmol/L.In summation,we have gotten the soluble expression production of hrpG gene in of E.coli M15 cells,which would provide a groundwork for the studies of the physiological activity and the specific binding site and the function regulation of HrpG protein. |
| |
| Keywords: | Xanthomonas campestris pv campestris HrpG Architecture characteristic Prokaryotic expression |
| 本文献已被 维普 万方数据 等数据库收录！ |
|
