鸭CYP7α1基因原核表达载体的构建及蛋白诱导表达  

Construction of Prokaryotic Expression Vector containing CYP7 alpha 1 Gene of Duck and the Expression of Induced Protein

作　　者：俞钦明[1,2] 甄霆[1] 陈昌义[2] 李欣钰[2] 陈阳[1] 张扬[1] 徐琪[1] 兰旅涛[2] 陈国宏[1] 

机构地区：[1]扬州大学动物科学与技术学院,江苏扬州225009 [2]江西农业大学动物科学与技术学院,江西南昌330045

出　　处：《中国家禽》2013年第20期6-10,共5页China Poultry

基　　金：江苏省科技支撑计划(BE2010391);现代农业产业技术体系建设专项资金(CARS-43-3)

摘　　要：本研究旨在构建鸭重组表达载体pET32a-CYP7α1,获取其融合蛋白,为开展鸭CYP7α1蛋白在脂类代谢中作用的深入研究提供基础。试验以樱桃谷鸭为材料,通过克隆获得CYP7α1基因的ORF序列,并连接至原核表达载体pET32a中,构建了重组表达载体pET32a-CYP7α1,转入表达宿主Rossetta-gam(iDE3)进行诱导表达。结果表明CYP7α1基因ORF区共编码512个氨基酸,SDS-PAGE电泳分析表明,重组蛋白大小约为59 ku,其主要以包涵体的形式存在,其最佳诱导条件为温度37℃,时间4 h,IPTG浓度0.6 mmol/L。This study was carried out to construct the duck recombinant expression vector to obtain the fusion protein to provide the basis for further researching the role of CYP7 alpha 1 protein in lipid metabolism. Taking Cherry Valley duck as the material in this study, the ORF of CYP7 alpha 1 gene of duck by cloning was inserted into pET32a prokaryotie expression vector. The recombinant plasmid named pET32a-CYP7a1 was constructed, and secondly transformed into Rossetta-gami（DE3）eells. SDS-PAGE analysis showed that the ORF region of CYP7 alpha 1 gene encodes 512 amino acidstotally, the recombinant protein weighed about 59 ku mainly existed in the form of inclusion body, and the optimum condition for induced exoression was 37℃ .4 h. 0.6 mmol/L IPTG

关 键 词：鸭 CYP7α1基因 原核表达 诱导表达 

分 类 号：S834[农业科学—畜牧学]