铜绿微囊藻气囊结构蛋白GvpC的表达纯化与晶体生长研究  

Expression,purification and crystallization of gas vesicle structure protein GvpC from the cyanobacterium Microcystis aeruginosa

作　　者：许柏英 张甜甜 Bo-ying XU;Tian-tian ZHANG(College of Life Sciences, Chongqing Normal University, Chongqing 401331, China;School of Life Sciences, University of Science and Technology of China, Heifei 230026, Anhui, China)

机构地区：[1]重庆师范大学生命科学学院,重庆401331 [2]中国科学技术大学生命科学学院,安徽合肥230026

出　　处：《山东大学学报（理学版）》2018年第11期1-8,共8页Journal of Shandong University(Natural Science)

基　　金：重庆市前沿与应用基础研究计划资助项目(cstc2015jcyjBX0142;cstc2016jcyjA0375)

摘　　要：对GvpC进行了初步晶体学研究。首先对GvpC进行分子克隆,利用原核表达系统在体外进行异源表达;结合变复性和凝胶过滤层析的方法纯化GvpC;通过圆二色谱的方法鉴定GvpC的复性效果;运用坐滴蒸汽扩散法进行晶体初筛。结果表明:GvpC在大肠杆菌表达系统中表达为包涵体;通过变复性和凝胶过滤层析能纯化出纯度较高的蛋白;圆二色谱证实复性后的GvpC形成了正确的二级结构,复性效果较佳;通过晶体初筛获得了GvpC的晶体,后续即可通过解析GvpC的晶体结构而获得其三维结构。Preliminary crystallographic study on the gas vesicle structure protein GvpC from the cyanobacterium Microcystis aeruginosa was performed. Firstly, gene gvpC was cloned and expressed; then, the recombinant protein was purified by denaturation and renaturation and gel filtration chromatography; the result of renaturation was detected by circular dichroism. Crystal screening was performed by sitting drop vapor diffusion techniques. GvpC was overexpressed as inclusion body in E. coli, and then the protein has been purified through denaturation and renaturation. The result of circular dichroism indicated that GvpC was refolded successfully in vitro. Crystals of GvpC were also obtained by crystal screening. Hence, the three-dimensional structure of GvpC could be determinated by solving its crystal structure.

关 键 词：铜绿微囊藻 气囊 GvpC 变复性 蛋白质晶体 

分 类 号：Q816[生物学—生物工程]