[1]侯绍华,柏丽华,郭晓宇,等.非洲猪瘟病毒p72蛋白杆状病毒表达系统的构建及其抗原性分析[J].中国预防兽医学报,2012,(01):33-36.[doi:10.3969/j.issn.1008-0589.2012.01.08]
 HOU Shao-hua,BAI Li-hua,GUO Xiao-yu,et al.Expression and identification of African swine fever virus protein p72 expressed in insect cells[J].Chinese journal of preventive veterinary medicine,2012,(01):33-36.[doi:10.3969/j.issn.1008-0589.2012.01.08]
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非洲猪瘟病毒p72蛋白杆状病毒表达系统的构建及其抗原性分析
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2012年01期
页码:
33-36
栏目:
病原生物学
出版日期:
2012-01-15

文章信息/Info

Title:
Expression and identification of African swine fever virus protein p72 expressed in insect cells
文章编号:
1008-0589(2012)01-0033-04
作者:
侯绍华柏丽华郭晓宇贾 红姜一曈朱鸿飞*
中国农业科学院 北京畜牧兽医研究所 动物医学研究室,北京 100193
Author(s):
HOU Shao-hua BAI Li-hua GUO Xiao-yu JIA Hong JIANG Yi-tong ZHU Hong-fei*
Beijing Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100091, China
关键词:
非洲猪瘟病毒p72蛋白杆状病毒表达抗原性
Keywords:
African swine fever virus p72 protein baculovirus expression antigenicity
分类号:
S852.65
DOI:
10.3969/j.issn.1008-0589.2012.01.08
文献标志码:
A
摘要:
为研究真核表达非洲猪瘟病毒(ASFV)主要结构蛋白p72,本研究从含ASFV p72基因全序列的重组质粒pGEX-6p-p72中扩增出1 941 bp的p72全长基因,将其插入于pFastBac HTa杆状病毒载体中,构建了重组质粒pFastBac HTa-p72,转化至感受态细胞DH10Bac中,获得重组杆粒rBacmid-p72。再将其转染至sf9昆虫细胞,获得重组杆状病毒。Western blot 和 夹心法ELISA分析表明ASFV p72基因在昆虫细胞sf9中获得了正确表达,重组p72蛋白可以被特异性抗ASFV血清、p72单克隆抗体识别,表明该蛋白特异性强、活性稳定,具有良好的抗原性。为进一步研究p72蛋白的结构、功能和免疫学特性奠定基础。
Abstract:
To express nucleocapsid p72 protein of African swine fever virus (ASFV), the p72 gene was amplified by PCR and cloned into pFastBac HTa vector. The resultant recombinant plasmid was transformed into DH10Bac competent cells to construct recombinant Bacmid-p72 which was transfected into Sf9 insect cells to generate recombinant baculovirus. The p27 protein was expressed in Sf9 cells and identified by western blot and sandwich ELISA. The recombinant p72 protein was recognized by anti-ASFV positive serum and p72 MAb, which could be used for ASFV detection method development.

参考文献/References:

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备注/Memo

备注/Memo:
收稿日期:2011-7-26
基金项目:国家十一五科技支撑计划项目(2009BADB4B00)
作者简介:侯绍华(1976-),男,河北昌黎人,助理研究员,博士,主要从事动物传染病及分子免疫学研究.
*通信作者:E-mail:bioclub@vip.sina.com
更新日期/Last Update: 2013-05-10