芜菁皱缩病毒P38蛋白的原核表达及抗血清的制备
Prokaryotic expression of P38 from Turnip Crinkle Virus and preparation of its polyclonal antibody
作者：张静(四川大学生命科学学院生物资源与生态环境教育部重点实验室)；朱峰(四川大学生命科学学院生物资源与生态环境教育部重点实验室)；陈应娟(四川大学生命科学学院生物资源与生态环境教育部重点实验室)；邓星光(四川大学生命科学学院生物资源与生态环境教育部重点实验室)；刘健(四川大学生命科学学院生物资源与生态环境教育部重点实验室)；林宏辉(四川大学生命科学学院生物资源与生态环境教育部重点实验室)；席德慧(四川大学生命科学学院生物资源与生态环境教育部重点实验室)
Author：ZHANG Jing(Key laboratory of Bio resources and Eco environment of Ministry of Education,College of Life Sciences, Sichuan University)；ZHU Feng(Key laboratory of Bio resources and Eco environment of Ministry of Education,College of Life Sciences, Sichuan University)；CHEN Ying-Juan(Key laboratory of Bio resources and Eco environment of Ministry of Education,College of Life Sciences, Sichuan University)；DENG Xing-Guang(Key laboratory of Bio resources and Eco environment of Ministry of Education,College of Life Sciences, Sichuan University)；LIU Jian(Key laboratory of Bio resources and Eco environment of Ministry of Education,College of Life Sciences, Sichuan University)；LIN Hong-Hui(Key laboratory of Bio resources and Eco environment of Ministry of Education,College of Life Sciences, Sichuan University)；XI De Hui(Key laboratory of Bio resources and Eco environment of Ministry of Education,College of Life Sciences, Sichuan University)
收稿日期：2013-12-26 年卷（期）页码：2015,52(2):441-445
期刊名称：四川大学学报: 自然科学版
Journal Name：Journal of Sichuan University (Natural Science Edition)
关键字：芜菁皱缩病毒; 外壳蛋白; 原核表达; 抗血清制备
Key words：TCV; Coat protein; Prokaryotic expression; Preparation of antiserum
基金项目：国家自然科学基金（31171835，31270290）; 四川省应用基础项目（2012JY0078）; 中央高校基本科研业务费（2011SCU04B34）
中文摘要
从感染芜菁皱缩病毒(TCV)的拟南芥中提取总RNA, 通过反转录PCR(RT PCR)扩增得到TCV的外壳蛋白基因P38. 在目的基因ORF框两边引入酶切位点 BamH Ⅰ和Sac Ⅰ, 并连接到表达载体pET30 a上, 导入大肠杆菌BL21(DE3)中, 37 ℃经0.5mM/L IPTG诱导6 h后获得了分子量约为48 kD的融合蛋白. 经Ni NTA纯化目的蛋白, 注射大白兔制备抗血清. Western blot结果表明制备好的抗血清能成功检测感染TCV的拟南芥.
英文摘要
The nucleotide sequence of the coat protein (P38) gene of Turnip crinkle virus was obtained using RT PCR from Arabidopsis thaliana infected with TCV. Restriction enzyme cutting sites of BamH Ⅰ and Sac Ⅰ were introduced to the ORF fragment of P38. Then the fragment was linked together with PET30 a and transformed into E.coli BL21(DE3). The target fusion protein(48 kD)was induced by 05mM/L IPTG at 37 ℃ for 6h. The expressed protein was purified by Ni NTA and anti P38 polyclonal antibody was prepared using rabbit. Western blot results showed that the antiserum could be used to detect the expression of P38 in infected Arabidopsis thaliana
【关闭】
