[1]王晓霞,郑亚东*,骆学农,等.绵羊痘病毒甘肃流行株糖蛋白基因ORF117 的克隆及其原核表达[J].中国预防兽医学报,2009,(05):337-341.
 WANG Xiao-xia,ZHENG Ya-dong*,LUO Xue-nong,et al.Cloning and prokaryotic expression of glycoprotein-encoding ORF117 from sheep pox virus Gansu strain[J].Chinese journal of preventive veterinary medicine,2009,(05):337-341.
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绵羊痘病毒甘肃流行株糖蛋白基因ORF117 的克隆及其原核表达
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2009年05期
页码:
337-341
栏目:
病毒及其病原生物学
出版日期:
2009-05-15

文章信息/Info

Title:
Cloning and prokaryotic expression of glycoprotein-encoding ORF117 from sheep pox virus Gansu strain
作者:
王晓霞12郑亚东1*骆学农1陈轶霞3李辉1侯俊玲1于三科2才学鹏1*
(1. 中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室/ 甘肃省动物寄生虫病重点实验室;
中国农业科学院人兽共患病重点实验室,甘肃兰州730046;2. 西北农林科技大学动物医学院,陕西杨陵712100;
3. 西北民族大学生命科学与工程学院,甘肃兰州730046)
Author(s):
WANG Xiao-xia12 ZHENG Ya-dong1* LUO Xue-nong1 CHEN Yi-xia3 LI Hui1 HOU Jun-ling1 YU San-ke2 CAI Xue-peng1*
(1. Key Laboratory of Zoonoses of CAAS, Key Laboratory of Veterinary Parasitology of Gansu Province,
State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, CAAS, Lanzhou 730046, China;2. College of Veterinary Medicine, Northwest Agriculture and Forestry University, Yangling 712100, China;
3. College of Life Science and Engineering, Northwest University of Nationalities, Lanzhou 730046, China)
关键词:
绵羊痘病毒ORF117 基因克隆原核表达
Keywords:
sheeppox virus ORF117 cloning prokaryotic expression
文献标志码:
A
摘要:
以甘肃景泰疑似羊痘(SP)患病绵羊的皮肤组织中提取的总DNA 为模板,经PCR 扩增获得SP 病毒(SPV) ORF117 基因。测序后分析表明,该基因由591 个核苷酸组成,编码196 个氨基酸;将该基因克隆于pET32载体中,构建重组表达质粒pETORF117,用IPTG 在不同条件下诱导表达,确定其最佳表达条件。SDS-PAGE 电泳分析表明,表达的融合蛋白分子质量为42 ku,主要以包涵体的形式存在。在IPTG 浓度为0.1 mM、温度为37 ℃、诱导6 h时表达量最大,约占菌体蛋白的30%  Western blot 试验表明,融合蛋白可被羊痘阳性血清识别,这为研究其在新型疫苗和诊断的研究奠定了基础。
Abstract:
 The glycoprotein-encoding ORF117 gene was cloned by PCR from genomic DNA of sheep pox virus isolated in Jingtai, Gansu Province. Sequence comparison showed that the ORF117 gene consist of a 591 bp open reading frame coding a 196  amino acid. The gene was subcloned into pET32 expression plasmid and expressed in by IPTG induction. SDS-PAGE analysis showed that the fusion protein had a molecular weight of 42 ku and mainly existed in form of inclusion body. The expressed protein accounted for more than 30 % of the total protein mass. Western blot confirmed that the fusion protein reacted to positive serum against sheep pox.

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更新日期/Last Update: 2009-06-05