| 抗菌肽V13KL原核表达载体的构建及其表达产物的鉴定 |
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| 引用本文： | 秦莹,乔木,关继羽,陈玥,孔德龙,安培培,唐博,陈育新,岳占碰,李子义.抗菌肽V13KL原核表达载体的构建及其表达产物的鉴定[J].兽医大学学报,2012(7):1043-1046. |
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| 作者姓名： | 秦莹 乔木 关继羽 陈玥 孔德龙 安培培 唐博 陈育新 岳占碰 李子义 |
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| 作者单位： | [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 |
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| 基金项目： | 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 |
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| 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Westernblotting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／LIPTG诱导6h便可检测到蛋白表达。
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| 关 键 词： | 抗菌肽 V13KL 原核表达 诱导条件 |
Construction of prokaryotic expression vector of antimicrobial peptides V13KL and the identification of its expression products |
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| Authors: | QIN Ying QIAO Mu GUAN Ji-yuI CHEN Yue KONG De-long AN Pei-pei TANG Bo CHEN Yu-xin YUE Zhan-peng LI Zi-yi |
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| Institution: | 1. J ilin Provincial Key Laboratory of Animal Embryo Engineering,College of Animal and Veterinary Medicine, Jilin University, Changchun 130062, China; 2. Dongling District Centre for Animal Epidemics Prevention and Csntrol , Shenyang 110015, China ; 3. Chang chun Proteli ght Pharmaceutical and Biotechnolog y Co. , Ltd , Changchun 130012 ,China) |
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| Abstract: | 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. |
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| Keywords: | antimicrobial peptide V13KL prokaryotic expression inducing conditions |
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