利用SUMO表达系统高效表达猪O型口蹄疫病毒VP0、VP1、VP3基因

›› 2013, Vol. 40 ›› Issue (9): 61-65.

利用SUMO表达系统高效表达猪O型口蹄疫病毒VP0、VP1、VP3基因

宋妮¹, 温永俊¹, 王凤雪¹, 武华^1,2

1.中国农业科学院特产研究所, 特种经济动物分子生物学国家重点实验室, 吉林长春 130112;
2. 华威特(北京)生物科技有限公司, 北京 100085

收稿日期:2013-02-25 出版日期:2013-09-20 发布日期:2013-09-18
通讯作者: 武华 E-mail:wuhua@sinovetah.com
作者简介:宋妮(1984-),女,山东人,硕士生,研究方向:动物疫苗与分子免疫学。
基金资助:
国家高技术研究发展计划(863计划)(2011AA10A213);吉林省特种经济动物疫病防控研究创新团队(20121823)。

Using SUMO Expression System to Efficiently Express VP0,VP1,VP3 Protein of Foot and Mouth Disease Virus Serotype O in Swine

SONG Ni¹, WEN Yong-jun¹, WANG Feng-xue¹, WU Hua^1,2

1. State Key Laboratory of Special Economic Animal Molecular Biology, Institute of Special Economic Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun 130112, China;
2. Sinovet (Beijing) Biotechnology Co., Ltd., Beijing 100085, China

Received:2013-02-25 Online:2013-09-20 Published:2013-09-18

摘要/Abstract

摘要： 根据GenBank中公布的猪O型口蹄疫病毒(foot and mouth disease virus,FMDV)全基因合成了FMDV结构蛋白前体蛋白P1基因,同时设计了扩增FMDV结构蛋白VP0、VP1和VP3基因的引物。以P1基因为模板,分别经PCR扩增获得FMDV VP0、VP1和VP3基因。扩增产物克隆于Blunt载体中,酶切后将目的片段连接到原核表达载体SUMO中,构建重组表达质粒SUMO-VP0、SUMO-VP1和SUMO-VP3,将重组质粒转化大肠杆菌BL21(DE3)plysS进行诱导表达。经SDS-PAGE电泳可见融合蛋白均获得高效表达,融合蛋白表观分子质量分别约为55、48和40 ku。在IPTG浓度为1.0 mmol/L,温度为37 ℃,诱导5 h时融合蛋白表达量最大。Western blotting结果表明,融合蛋白均可被FMDV阳性血清识别,反应原性良好。

关键词: 猪O型口蹄疫病毒; VP0、VP1、VP3基因; SUMO; 高效表达

Abstract: Structural protein precursor P1 gene of foot and mouth disease virus (FMDV) serotype O in swine was synthesised according to published sequences in GenBank.The structural protein VP0,VP1 and VP3 gene primers were designed depending on these specific sequences,and were obtained by PCR method according to the P1 gene, respectively.The PCR products were cloned into Blunt vector,and were digested and ligated into the prokaryotic expression vector SUMO.These recombinant expression plasmids,SUMO-VP0, SUMO-VP1 and SUMO-VP3,were transformed into E.coli BL21(DE3)plysS.For detecting the efficient expression,these fusion protein products expressed by the E.coli were checked by SDS-PAGE electrophoresis,and molecular weight were 55,48 and 40 ku,respectively.The largest ratio of the bacterial protein was induced under the conditions of IPTG concentration of 1.0 mmol/L,temperature of 37 ℃ and last for 5 h. The results of Western blotting showed that the fusion proteins had a good reaction with the FMDV positive serum.

Key words: FMDV serotype O in swine; VP0,VP1,VP3 gene; SUMO; high expression

中图分类号:

Q786

宋妮, 温永俊, 王凤雪, 武华. 利用SUMO表达系统高效表达猪O型口蹄疫病毒VP0、VP1、VP3基因[J]. , 2013, 40(9): 61-65.

SONG Ni, WEN Yong-jun, WANG Feng-xue, WU Hua. Using SUMO Expression System to Efficiently Express VP0,VP1,VP3 Protein of Foot and Mouth Disease Virus Serotype O in Swine[J]. China Animal Husbandry & Veterinary Medicine, 2013, 40(9): 61-65.

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