猪干扰素-δ基因原核表达载体的构建及高效表达

doi:10.16431/j.cnki.1671-7236.2016.04.013

›› 2016, Vol. 43 ›› Issue (4): 940-945.doi: 10.16431/j.cnki.1671-7236.2016.04.013

猪干扰素-δ基因原核表达载体的构建及高效表达

李红¹, 刘成倩¹, 严华祥¹, 王家豪², 易建中¹

1. 上海市农业科学院畜牧兽医研究所, 上海 201106;
2. 上海农林职业技术学院, 上海 201699

收稿日期:2015-10-08 出版日期:2016-04-20 发布日期:2016-04-27
通讯作者: 易建中 E-mail:yijianzhong@yahoo.com
作者简介:李红(1983-),女,山东烟台人,博士,助理研究员,研究方向:动物分子生物学和免疫学,Tel:021-62204612;E-mail:lihong20061029@163.com
基金资助:
上海市科技人才计划(14YF1414600)

Construction and High Level Expression of Prokaryotic Expression Vector of Pig Interferon-δ Gene

LI Hong¹, LIU Cheng-qian¹, YAN Hua-xiang¹, WANG Jia-hao², YI Jian-zhong¹

1. Institute of Animal Husbandry & Veterinary Science, Shanghai Academy of Agricultural Sciences, Shanghai 201106, China;
2. Shanghai Vocational College of Agriculture and Forestry, Shanghai 201699, China

Received:2015-10-08 Online:2016-04-20 Published:2016-04-27

摘要/Abstract

摘要： 干扰素(interferon,IFN)因具有广谱抗病毒、抗肿瘤活性及免疫调节作用而备受关注。本研究根据GenBank上公布的猪IFN-δ核苷酸序列设计1对引物,通过PCR方法扩增出猪 IFN-δ基因,片段大小为513 bp,并将其克隆到原核表达载体上得到重组质粒。将重组质粒转化大肠杆菌Transetta BL21(DE3)受体菌,经SDS-PAGE电泳分析,结果表明,pET-30a-DsbA-IFN-δ表达量较高,表达的融合蛋白分子质量约20 ku,且主要以包涵体形成存在,经Western blotting分析发现表达的融合蛋白能与His标签单克隆抗体发生特异性反应。

关键词: 猪; 干扰素-δ; 原核表达

Abstract: Interferon (IFN) had recieved much more attention because of its broad-spectrum antiviral, antitumor activity and immune regulation.One pair of primers were designed for amplifying pig IFN-δ gene with PCR method according to the relevant sequence from GenBank.The IFN-δ gene was cloned into prokaryotic expression vector, and the recombinant plasmid was obtained.We transformed the recombinant plasmid into E.coli Transetta BL21(DE3) strain and the protein expression was identified by SDS-PAGE and Western blotting.The results showed that pET-30a-DsbA-IFN-δ recombinant protein was 20 ku and expressed mainly in the form of inclusion body, these expressed protein could specific react with His-tag monoclonal antibody.

Key words: pig; interferon-δ; prokaryotic expression

中图分类号:

S852.2

李红, 刘成倩, 严华祥, 王家豪, 易建中. 猪干扰素-δ基因原核表达载体的构建及高效表达[J]. , 2016, 43(4): 940-945.

LI Hong, LIU Cheng-qian, YAN Hua-xiang, WANG Jia-hao, YI Jian-zhong. Construction and High Level Expression of Prokaryotic Expression Vector of Pig Interferon-δ Gene[J]. , 2016, 43(4): 940-945.

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猪干扰素-δ基因原核表达载体的构建及高效表达

Construction and High Level Expression of Prokaryotic Expression Vector of Pig Interferon-δ Gene

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