犬瘟热病毒V蛋白的表达、鉴定及亚细胞定位

doi:10.16431/j.cnki.1671-7236.2016.07.010

《中国畜牧兽医》---唯一指定的官方网站 ›› 2016, Vol. 43 ›› Issue (7): 1723-1728.doi: 10.16431/j.cnki.1671-7236.2016.07.010

犬瘟热病毒V蛋白的表达、鉴定及亚细胞定位

刘莉¹, 郝福星¹, 袁维峰², 封家旺³

1. 江苏农牧科技职业学院, 泰州 225300;
2. 中国农业科学院北京畜牧兽医研究所, 北京 100193;
3. 西藏职业技术学院, 拉萨 850030

修回日期:2016-02-23 出版日期:2016-07-20 发布日期:2016-07-22
通讯作者: 封家旺 E-mail:fengjiawang126@126.com
作者简介:刘莉(1981-),女,江苏铜山人,博士,讲师,主要从事动物医学专业研究与教学工作,E-mail:jsahvcll@163.com;郝福星(1982-),男,江苏如皋人,硕士,讲师,主要从事动物医学专业研究与教学工作,E-mail:vethfx@163.com
基金资助:
公益性行业（农业）科研专项（201303042）

Expression, Identification and Subcellular Localization of V Protein of Canine Distemper Virus

LIU Li¹, HAO Fu-xing¹, YUAN Wei-feng², FENG Jia-wang³

1. Jiangsu Animal Husbandry and Veterinary College, Taizhou 225300, China;
2. Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China;
3. Tibet Vocational Technical College, Lhasa 850030, China

Revised:2016-02-23 Online:2016-07-20 Published:2016-07-22

摘要/Abstract

摘要：

为研究犬瘟热病毒（canine distemper virus，CDV） V蛋白的功能，将CDV V基因片段与pGEX-6P-1载体连接，构建pGEX-6P-1-CDV-V重组表达质粒，通过原核表达系统表达了V蛋白，并将纯化的V蛋白免疫BALB/c小鼠，制备阳性血清。同时，将CDV V基因片段与pcDNA3.1载体连接，构建pcDNA3.1-CDV-V重组表达质粒，经转染Vero细胞后，用激光共聚焦技术确定V蛋白在真核细胞中的分布及亚细胞定位。结果显示，成功表达了V蛋白，并制备了阳性血清。重组质粒pcDNA3.1-CDV-V在外源真核细胞Vero中获得了表达，表达蛋白主要聚集于细胞质。本试验结果为进行CDV V蛋白的功能研究奠定了基础。

关键词: 犬瘟热病毒; V蛋白; 原核细胞; 亚细胞定位

Abstract:

In order to study the function of canine distemper virus (CDV) V protein,V gene fragment was inserted into the pGEX-6p-1 vector,and then pGEX-6p-1-CDV-V recombinant expression plasmid was obtained.Purified V protein immunized BALB/c mice to prepare the positive serum.Meanwhile,to confirm the distribution of V protein and subcellular localization with confocal laser technology,a eukaryotic expression recombinant plasmid pcDNA3.1-CDV-V was built,then transfected into Vero cells.The results showed that V protein was successfully expressed and the positive serum was prepared.The recombinant plasmid pcDNA3.1-CDV-V was expressed in Vero cells and mainly expressed in cytoplasm.The results laid a function for functional studies of CDV V protein.

Key words: canine distemper virus; V protein; prokaryotic cell; subcellular localization

中图分类号:

S852.65⁺5

刘莉, 郝福星, 袁维峰, 封家旺. 犬瘟热病毒V蛋白的表达、鉴定及亚细胞定位[J]. 《中国畜牧兽医》---唯一指定的官方网站, 2016, 43(7): 1723-1728.

LIU Li, HAO Fu-xing, YUAN Wei-feng, FENG Jia-wang. Expression, Identification and Subcellular Localization of V Protein of Canine Distemper Virus[J]. , 2016, 43(7): 1723-1728.

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犬瘟热病毒V蛋白的表达、鉴定及亚细胞定位

Expression, Identification and Subcellular Localization of V Protein of Canine Distemper Virus

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