[1]高 明,魏 华*.重组猪IFN-α原核表达及其活性分析 [J].中国预防兽医学报,2009,(11):902-905.
 GAO Ming,WEI Hua*.Expression of recombinant porcine alpha-interferon in E.coli and the anti-virus activity analysis[J].Chinese journal of preventive veterinary medicine,2009,(11):902-905.
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重组猪IFN-α原核表达及其活性分析


 

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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2009年11期
页码:
902-905
栏目:
免疫学
出版日期:
2009-11-15

文章信息/Info

Title:
Expression of recombinant porcine alpha-interferon in E.coli and the anti-virus activity analysis


作者:
高 明魏 华*
(沈阳农业大学 畜牧兽医学院,辽宁 沈阳 110161)
Author(s):
GAO Ming WEI Hua*
(College of Animal Husbandry and Veterinary, Shenyang Agricultural University, Shenyang 110161, China)
关键词:
重组α干扰素原核表达
Keywords:
 porcine recombinant alpha-interferon prokaryotic expression
文献标志码:
A
摘要:
为了表达猪重组α干扰素(rIFN-α)并对它的生物学特性进行研究,本研究采用RT-PCR从猪脾淋巴细胞中扩增出猪IFN-α基因,以pPROExHTa为载体构建了表达重组质粒poIFN-α,转化宿主菌BL21,经IPTG诱导猪rIFN-α获得了表达,其分子量约22 ku。该重组蛋白以包涵体形式存在,其表达量占总菌体蛋白7 %。采用Ni-Ni金属螯合亲和层析方法纯化,其纯度达到80 %以上,包涵体经复性后,在WISH/VSV系统上,采用细胞病变抑制法测定表明,其稀释度在小于1∶103时才有抗病毒活性。本实验表达的猪rIFN-α具有一定的抗病毒活性,为研究具有广谱抗病毒效应的猪干扰素具有重要意义。
Abstract:
Porcine alpha interferon (IFN-α) gene was amplified by using RT-PCR from pig spleen lymphocytes and subcloned into pPROExHTa expression vector, transformed into host bacteria BL21. Recombinant porcine IFN-αwas about 22 ku expressed by IPTG-induction, accounted for up to 7 % of total bacterial protein. Porcine rIFN-α achieved purity to above 80 % by Ni-Ni metal chelate affinity chromatography purification, The rIFN-α possessed a anti-vesicular stomatitis virus activity at 2 900 IU/mg.

参考文献/References:

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备注/Memo

备注/Memo:
收稿日期:2009-04-18
基金项目:兽医生物技术国家重点实验室开放课题基金(SKLVBF200807)
作者简介:高 明(1970-),男,辽宁阜新人,讲师,主要从事基础兽医学研究.
*通信作者:E-mail:gaomingjiepou@163.com
更新日期/Last Update: 2009-12-24