蓖麻毒素A链在麦胚无细胞系统中的表达  

Expression of RTA by wheat germ cell-free system

作　　者：肖世峰 许娜[2] 姚林芳[3] 孙成彪 刘洋[5] 黄燕霞[1] 李松岩[2] 万家余[1] 王兴龙[1] 刘文森[1] 

机构地区：[1]军事医学科学院军事兽医研究所/吉林省人兽共患病防控重点实验室,吉林长春130122 [2]吉林医药学院,吉林吉林市132013 [3]吉林农业大学生命科学院,吉林长春130118 [4]吉林农业大学食品科学与工程学学院,吉林长春130118 [5]吉林大学白求恩医学院药理学系,吉林长春130021

出　　处：《中国兽医学报》2015年第11期1804-1808,共5页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(项目编号31372487);中国博士后科学基金资助项目(2014M552637)

摘　　要：本试验利用麦胚无细胞系统表达蓖麻毒素A链(RTA):通过PCR方法扩增蓖麻毒素A链基因序列并连接至pMD18-T载体。将测序正确的RTA序列克隆到Flexi表达载体,获得Flexi-RTA重组质粒。将该重组质粒加入到麦胚无细胞表达系统中进行表达。表达产物经SDS-PAGE电泳分析相对分子质量约为32 000,通过Western blot、ELISA实验鉴定为RTA。本方法表达的RTA在蓖麻毒素诊断、疫苗研发、抗癌药物研制等方面具有广阔应用前景。This experiment is about expressing ricin A chain（RTA）by wheat germ cell-free system.RTA gene sequence was obtained by PCR amplification.The RTA gene sequence was then linked to the pMD18-T vector.After identified correct,the RTA gene sequence was cloned to Flexi expression vector to build Flexi-RTA recombinant plasmid.The Flexi-RTA recombinant plasmid was then added into wheat germ cell-free system to express proteins.The expressed protein which molecular weight is 32 000,was identified as RTA by SDS-PAGE,Western blot and ELISA.The expressed RTA has multiple-use in ricin poisoning diagnosis,vaccine research and development and Anti-cancer drug as well.

关 键 词：蓖麻毒素A链 Flexi载体 麦胚无细胞表达系统 

分 类 号：Q786[生物学—分子生物学]