[1]曹宗喜,潘全会,齐 岩,等.PRRSV变异株ORF6基因重组杆状病毒的构建及其在昆虫细胞的表达[J].中国预防兽医学报,2009,(04):261-265.
 CAO Zong-xi,PAN Quan-hui,QI Yan,et al.Construction of recombinant baculovirus expressing ORF6 gene of PRRSV[J].Chinese journal of preventive veterinary medicine,2009,(04):261-265.
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PRRSV变异株ORF6基因重组杆状病毒的构建及其在昆虫细胞的表达
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2009年04期
页码:
261-265
栏目:
病毒及分子生物学
出版日期:
2009-04-15

文章信息/Info

Title:
Construction of recombinant baculovirus expressing ORF6 gene of PRRSV
作者:
曹宗喜12潘全会3齐 岩12孔留五12何逸民12罗玉均12秦宏阳12张翔峰12张得玉12张桂红12*
(1. 华南农业大学 兽医学院,广东 广州 510642;2. 农业部动物疫病防控重点开放实验室,广东 广州 510642;
3. 美国礼来(亚洲)公司动物保健处,上海 200021)
Author(s):
CAO Zong-xi12 PAN Quan-hui3 QI Yan12 KONG Liu-wu12 HE Yi-min12 LUO Yu-jun12 QIN Hong-yang12 ZHANG Xiang-feng12 ZHANG De-yu12 ZHANG Gui-hong12*
(1. College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China; 2. Key Laboratory of Animal Disease Control and Prevention, Ministry of Agriculture, Guangzhou 510642, China; 3. Elanco Animal Health A Division of Eli Lilly Asia, Inc., Shanghai 200021, China)
关键词:
PRRSV杆状病毒表达克隆
Keywords:
porcine reproductive and respiratory syndrome virus (PRRSV) baculovims expression c1oning
文献标志码:
A
摘要:
参照GenBank中登录的猪繁殖与呼吸综合征病毒(PRRSV) XH-GD株ORF6 (M蛋白)基因序列,设计合成了一对引物,扩增PRRSV ORF6基因。将该基因克隆于转移质粒载体pFcDNA3.1 (+)中,获得重组转移质粒pFcDNA3.1-ORF6并将其转化DH10Bac细胞,得到重组穿梭质粒Bacmid-ORF6,再将其转染昆虫细胞Sf9,获得重组杆状病毒rBacmid-ORF6,经PCR鉴定证实目的基因正确地插入到杆状病毒基因组的CMV启动子下游;经过间接免疫荧光试验(IFA)检测,M蛋白在Sf9细胞中得到了表达,而且表达的产物具有特异免疫学反应性。
Abstract:
The ORF6 (M protein) gene of PRRSV was amplified by PCR using a pair of primers based on the ORF6 sequences of a mutant PRRSV. The PCR product was cloned into the transfer vector pFcDNA3.1 (+) and transformed into DH10Bac-host cells to produce recombinant shuttle plasmids Bacmid-ORF6, which was then used to transfect Sf9 insect cells. Indirect flurescent assay (IFA) confirmed that the ORF6 gene was expressed in Sf9 cells. The expression product could specially react to the sera of PRRSV.

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备注/Memo

备注/Memo:
收稿日期:2009-02-10
基金项目:863计划(2006AA10A204);国家生猪现代农业产业技术体系;广东省农业类计划项目
(2007A020300006-4)
作者简介:曹宗喜(1982-),男,河南新乡人,博士研究生,主要从事病毒的分子生物学和免疫学方面的研究;
潘全会(1979-),男,河南确山人,硕士,主要从事临床及分子生物学和免疫学方面的研究.
*通信作者:E-mail:guihongzh@scau.edu.cn
更新日期/Last Update: 2009-06-09