抗菌肽V13KL原核表达载体的构建及其表达产物的鉴定  

Construction of prokaryotic expression vector of antimicrobial peptides V13KL and the identification of its expression products

作　　者：秦莹[1] 乔木[2] 关继羽[1] 陈玥[1] 孔德龙[1] 安培培[1] 唐博[1] 陈育新 岳占碰[1] 李子义[1] 

机构地区：[1]吉林大学畜牧兽医学院动物胚胎工程吉林省重点实验室,吉林长春130062 [2]沈阳市东陵区动物疫病预防控制中心,辽宁沈阳110015 [3]长春普莱医药生物技术有限公司,吉林长春130012

出　　处：《中国兽医学报》2012年第7期1043-1046,共4页Chinese Journal of Veterinary Science

基　　金：国家转基因生物新品种培育科技重大专项(2011ZX08008-005);国务院华侨办华人创业团队基金重点专项资助项目

摘　　要：将V13KL编码基因片段克隆到原核表达载体pET30b(+),并在目的蛋白N端设计肠激酶酶切位点,构建出pET30b(+)-VK13L重组质粒,转化大肠杆菌表达菌株BL21(DE3),优化诱导条件,使得抗菌肽得到了高效表达。经Tricine-SDS-PAGE和Western blotting鉴定,目的蛋白的相对分子质量与预期一致。结果表明,V13KL基因成功克隆并且获得表达,在诱导体系中加入1mmol/L IPTG诱导6h便可检测到蛋白表达。V13KL is a kind of antimicrobial peptide,which is obtained by chemical synthesis. It is with characteristic of highly efficient, broad-spectrum and low hemolysis. First, V13KL encoding gene fragment was cloned into the pro- karyotic expression vector pET30b （＋）, and enterokinase restriction sites was designed at N-terminal. Then, pET30b（＋）-VK13L recombinant plasmid was transformed into E. coli expression strain BL21 （DE3）. The induction conditions were optimized, which make the peptide highly expressed. The identification by Tricine-SDS-PAGE and Western blotting indicated that the molecular weight of the protein was as expected. The results showed that V13KL gene was successfully cloned and expressed , and the protein expression could be detected in the induction system when 1 mmol/L IPTG was added up to 6 h.

关 键 词：抗菌肽 V13KL 原核表达 诱导条件 

分 类 号：Q78[生物学—分子生物学] S852.2[农业科学—基础兽医学]