梅花鹿朊蛋白核心片段的基因克隆与高效表达  

Cloning and High-level Expression of a Core Fragment Gene of Prion Protein from Sika Deer(Cervus nippon)

作　　者：刘东[1] 卢士英[1] 李岩松[1,2] 李兆辉[1] 王光明[1] 张媛媛[1] 周玉[1] 宋杰[1] 柳增善[1] 任洪林[1] 

机构地区：[1]人兽共患病研究教育部重点实验室吉林大学人兽共患病研究所畜牧兽医学院,吉林长春130062 [2]吉林大学军需科技学院,吉林长春130062

出　　处：《中国畜牧兽医》2012年第3期78-80,共3页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金项目(30871956);吉林大学农学部博士科研启动基金项目(4305050102C9)

摘　　要：本试验根据梅花鹿朊蛋白基因序列设计引物,利用PCR的方法从梅花鹿基因组DNA中扩增朊病毒蛋白酶抵抗区域PrPres,将该片段分别与表达载体pET-Trx和pET-His连接,构建重组表达载体pET-Trx-PrPres和pET-His-PrPres。分别将两个重组表达载体转入E.coli BL21(DE3)plys宿主菌中,37℃诱导4h,经SDS-PAGE分析,Trx-PrPres和His-PrPres表达量分别为38.2%和30.1%。The gene of prion protein protease resistance fragment PrPres of the Sika deer was amplified by PCR using a pair of specific primers.The product of PCR was cloned into the expression vectors of pET-Trx and pET-His.The restructuring plasmids were transformed into the host bacterium of E.coli BL21（DE3） plys,at 37 ℃ for 4 h.SDS-PAGE result illustrated that fusion proteins of Trx-PrPres and His-PrPres were highly expressed.The expression level of fusion proteins were 38.2% and 30.1%,respectively.

关 键 词：梅花鹿 朊蛋白 朊病毒 蛋白酶抗性 原核表达 

分 类 号：Q78[生物学—分子生物学]