北京油鸡β-防御素-7基因原核表达质粒的构建及表达  

Construction and prokaryotic expression of recombinant plasmid containing β-gallinacin-7 gene of Beijing fatty chicken

作　　者：朱超[1,2] 冯宝刚[1,2] 陈献伟[2] 张明[1] 关伟军[2] 

机构地区：[1]广西大学动物科学技术学院,广西南宁530004 [2]中国农业科学院北京畜牧兽医研究所,北京100193

出　　处：《黑龙江畜牧兽医》2010年第4期12-15,共4页Heilongjiang Animal Science And veterinary Medicine

基　　金："十一五"国家科技支撑计划项目(2008BADB2B01;2006BAD13B08);广西科技厅科技攻关项目(桂科攻0815008-2-4);广西大学研究生创新项目(2008105930905D001)

摘　　要：为了研究β-防御素-7基因在大肠杆菌中的原核表达,试验以pGEM-T-GAL-7为模板,经PCR扩增GAL-7基因的编码序列,克隆入原核表达载体pGEX-4T-1,转化E.coliDH5α并在E.coliBL21(DE3)中诱导表达,并对目的蛋白进行SDS-PAGE电泳和Western-blot鉴定。结果表明:重组质粒pGEX-4T-1-GAL-7经PCR及酶切测序鉴定,证明质粒构建正确;重组质粒经诱导表达后,可表达出分子质量约为33ku的GAL-7蛋白;表达产物以融合蛋白形式存在,在37℃条件下诱导4h表达量最高,最佳的诱导浓度为0.1mmol/L。In order to investigate the recombinant plasmid for prokaryotie expression of β - Gallinaein -7 gene in E. coli DHSa, the encoding sequence of GAL - 7 was amplified by PCR using the pGEM - T - GAL - 7 and inserted into prokaryotie expression vector pGEX -4T - 1. The constructed recombinant plasmid was transformed to E. coli DH5ct and expressed in the E. coli BL21 （ DE3 ）. The expressed product was identi- fied by SDS - PAGE and Western - blot. The results showed that the recombinant plasmid pGEX - 4T - 1 - GAL - 7 was correctly constructed by PCR method and restriction enzyme analysis and sequence. The GAL - 7 protein, with a relative molecular mass of about 33 ku, was expressed in a form of fusion protein. The expressed protein reached the maximum after 4 hours in 37℃ at 0. 1 mmol/L.

关 键 词：Β-防御素 克隆 原核表达 

分 类 号：S831.89[农业科学—畜牧学]