中国农业科学院农业信息研究所 中国农学会
Agricultural Information Institute of CAAS China Association of Agricultural Science Societies
En
Ch
中国农业科学院农业信息研究所 中国农学会 主办

AI检索

订阅

返回顶部
当前位置: 首页 > 文章 > V5标记的猪圆环病毒2型Cap蛋白重组杆状病毒表达载体的构建 中国畜牧兽医 2018 (7)
Position: Home > Articles > V5标记的猪圆环病毒2型Cap蛋白重组杆状病毒表达载体的构建 China Animal Husbandry & Veterinary Medicine 2018 (7)

V5标记的猪圆环病毒2型Cap蛋白重组杆状病毒表达载体的构建

作  者：
徐国;梁海英;曾智勇;王彬;黄涛;汤德元;代振江;叶百川;张爱琼;何小莉;咸文
关键词：
猪圆环病毒2型(PCV2);Cap-V5蛋白;重组杆状病毒;表达载体
摘  要：
试验旨在利用真核表达系统构建V5标记的猪圆环病毒2型(PCV2)Cap蛋白的表达载体.采用基因工程技术将V5标签引入到PCV2 ORF2基因C末端,并将标记基因定向克隆入pFastBacHTA载体,将pFastBacHTA-ORF2-V5重组转移载体转化大肠杆菌DH10Bac感受态细胞,使用脂质体介导法将鉴定后的重组杆状病毒质粒转染于Sf9细胞中;运用间接免疫荧光试验(IFA)、SDS-PAGE和Western blotting试验对Sf9细胞中V5标记Cap蛋白的表达进行验证.结果显示,本试验成功将V5标签引入到PCV2 ORF2基因末端,pFastBacHTAORF2-V5重组转移载体转化大肠杆菌DH10Bac感受态细胞后获得了Bacmid-ORF2-V5重组杆状病毒质粒;IFA、SDS-PAGE和Western blotting试验结果显示,在Sf9细胞中能检测到重组杆状病毒V5标记的PCV2Cap蛋白,具有良好的反应原性,说明本试验成功获得一株rAcMNPV Cap-V5重组杆状病毒.试验结果为PCV2标记亚单位疫苗的研制提供一定的理论依据.
作  者：
XU Guo;LIANG Haiying;ZENG Zhiyong;WANG Bin;HUANG Tao;TANG Deyuan;DAI Zhenjiang;YE Baichuan;ZHANG Aiqiong;HE Xiaoli;XIAN Wen;College of Animal Science,Guizhou University;
单  位：
XU Guo%LIANG Haiying%ZENG Zhiyong%WANG Bin%HUANG Tao%TANG Deyuan%DAI Zhenjiang%YE Baichuan%ZHANG Aiqiong%HE Xiaoli%XIAN Wen%College of Animal Science,Guizhou University
关键词：
porcine circovirus type 2(PCV2);;Cap-V5 protein;;recombinant baculovirus;;expression vector
摘  要：
The aim of the experiment was to construct the expression vector of V5-labeled porcine circovirus type 2(PCV2)Cap protein using eukaryotic expression system.The V5 tag was introduced into the end of the PCV2 ORF2 gene using genetic engineering,and the tagged gene was cloned into the pFastBacHTA vector.Recombinant transfer vector pFastBacHTA-ORF2-V5 was transformed into E.coli DH10 Bac.The identified recombinant bacmid was transfected into Sf9 cells using liposome-mediated method;Indirect immunofluorescence assay(IFA),SDS-PAGE and Western blotting were used to test the expression of V5-labeled Cap protein in Sf9 cells.The results showed that this experiment successfully introduced the V5 tag to the end of the PCV2 ORF2 gene.After the recombinant transfer vector pFastBacHTA-ORF2-V5 transformed into E.coli DH10 Bac,Bacmid-ORF2-V5 recombinant bacmid was obtained.The result of IFA,SDSPAGE and Western blotting showed that it could be expressed in Sf9 cells.The recombinant baculovirus V5-labeled PCV2 Cap protein was detected to have good reactogenicity,indicating that this experiment successfully obtained a rAcMNPV-Cap-V5 recombinant baculovirus.The test resultsprovided a theoretical basis for the development of PCV2 marker subunit vaccine.

相似文章

导出引用
计量
文章访问数: 15
HTML全文浏览量: 0
PDF下载量: 0

所属期刊

推荐期刊

友情链接Friendly link
农业信息研究所Agricultural information institute 农业智能知识服务平台Agricultural intelligent knowledge service platform 农业学术服务平台Agricultural Knowledge Service System 中国知网CNKI 维普网VIP 万方WANFANG DATA
联系我们Contact us

电话Tell：010-82109652-808

邮箱E-mail：agrijournal@caas.cn

服务Service
关于我们About us 意见建议Opinions and suggest
版权说明Copyright
单位：中国农业科学院农业信息研究所 中国农学会 © 2017中国农业科学院农业信息研究所 ICP备案号：京ICP备09089781号-46