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铜绿微囊藻气囊结构蛋白GvpC的表达纯化与晶体生长研究
引用本文:许柏英,张甜甜.铜绿微囊藻气囊结构蛋白GvpC的表达纯化与晶体生长研究[J].山东大学学报(理学版),2018,53(11):1-8.
作者姓名:许柏英  张甜甜
作者单位: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
基金项目:重庆市前沿与应用基础研究计划资助项目(cstc2015jcyjBX0142);重庆市前沿与应用基础研究计划资助项目(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。

关 键 词:铜绿微囊藻  气囊  GvpC  变复性  蛋白质晶体  
收稿时间:2018-04-17

Expression,purification and crystallization of gas vesicle structure protein GvpC from the cyanobacterium Microcystis aeruginosa
Bo-ying XU,Tian-tian ZHANG.Expression,purification and crystallization of gas vesicle structure protein GvpC from the cyanobacterium Microcystis aeruginosa[J].Journal of Shandong University,2018,53(11):1-8.
Authors:Bo-ying XU  Tian-tian ZHANG
Institution:1. College of Life Sciences, Chongqing Normal University, Chongqing 401331, China
2. School of Life Sciences, University of Science and Technology of China, Heifei 230026, Anhui, China
Abstract: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.
Keywords:Microcystis aeruginosa  gas vesicle  GvpC  denaturation and renaturation  protein crystal  
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