乳酸菌素基因PlnE和PlnF在大肠杆菌中的异源表达  

Heterologly Expression of Plantaricin Genes PlnE and PlnF in E.coli

作　　者：杨芳[1] 余占桥[1] 张日俊[1] 

机构地区：[1]中国农业大学饲料生物技术实验室,动物营养学国家重点实验室,北京100193

出　　处：《中国畜牧兽医》2010年第12期55-59,共5页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金“高产广谱细菌素工程菌的构建表达和特性研究”(30972124);国家高技术研究发展计划(863计划)重大项目“生物兽药新产品研究与创制”(2006AA10A208)

摘　　要：为获得高产量纯品植物乳杆菌素PlnE和PlnF,研究其开发生物兽药的可能,试验通过人工合成植物乳杆菌素基因PlnE和PlnF,构建表达载体pGEX-4T-E和pGEX-4T-F,经IPTG诱导表达条件优化后,进行亲和层析纯化融合表达蛋白,并使用SDS-PAGE进行鉴定。试验结果表明,pGEX-4T-E和pGEX-4T-F构建成功,纯化后获得2种产量高达30%的纯品融合蛋白,SDS-PAGE鉴定两融合蛋白表达正确。说明PlnE和PlnF基因片段编码的多肽能在原核细胞中正确表达,为进一步研究该细菌素的生物活性奠定了基础。In order to obtain high-yield pure plantaricin,and research on biological veterinary,plantaricin PlnE and PlnF were synthesised by Invitrogen.The aim genes were inserted into a bacterial expression vector pGEX-4T-1 to construct two prokaryotic expression vectors of pGEX-4T-E and pGEX-4T-F.After IPTG induction and express optimization,the fusion proteins were purified by affinity chromatography and detected by SDS-PAGE.The results showed that expression vectors were constructed correctly,more than 30% pure fusion proteins were obtained,which indicated peptides PlnE and PlnF could be accurately expressed in prokaryote cells,and lay a foundation for further researches on their bioactivity.

关 键 词：植物乳酸杆菌 细菌素 原核表达 融合蛋白 

分 类 号：Q786[生物学—分子生物学]