|
|
|
猪瘟病毒E2 基因和牛疱疹病毒Ⅰ型
UL49基因的共表达
|
|
|
|
刘燕,田志军,郑宝亮,周伦江,周艳君,仇华吉,童光志
|
|
|
|
（中国农业科学院 哈尔滨兽医研究所 兽医生物技术国家重点实验室，黑龙江 哈尔滨150001）
|
|
|
|
摘要：
|
|
为了探讨牛疱疹病毒Ⅰ型(BHV-1)UL49基因编码的膜蛋白VP22对猪瘟病毒E2基因表达水平的影响，构建了单独表达E2蛋白的pcDNA3.1-E2及融合表达E2-VP22的pcDNA3.1-E2-UL49。将pcDNA3.1-E2与pcDNA3.1-E2-UL49分别转染293 T细胞，间接免疫荧光试验结果显示，单独表达E2蛋白的细胞，其荧光主要固缩于核周，而融合表达E2-VP22蛋白的细胞在核周及细胞质内都有大量的荧光；Western-blot分析结果显示，VP22蛋白与E2蛋白都获得了表达，但融合了VP22蛋白的E2蛋白表达量要相对高一些（P<0.01），表明VP22蛋白可以促进E2蛋白的表达。 |
|
关键词：
|
|
猪瘟病毒；E2基因；UL49基因；体外表达 |
|
中图分类号：S 852.659.6：Q 786文献标识码：
A 文章编号：1673-4696(2006)10-0777-05 |
|
|
Co-expression of classical swine fever virus E2 gene with
bovine herpesvirus 1 UL49 gene |
|
|
|
LIU Yan,TIAN Zhi-jun,ZHENG Bao-liang,ZHOU Lun-jiang,
ZHOU Yan-jun,QIU Hua-ji,TONG Guang-zhi |
|
|
|
(National Key Laboratory of Veterinary Biotechnology/Harbin Veterinary Research Institute,Chinese Academy
of Agricultural Sciences,Harbin 150001,China) |
|
|
Abstract: n order to investigate the influence of VP22 protein on expression of E2 gene in vitro，two recombinant eukaryotic expression vectors, pcDNA3.1-E2 and pcDNA3.1-E2-UL49, were constructed by inserting E2 gene and E2-VP22 protein gene into pcDNA3.1(+), respectively. The expressed products in 293 T cells transfected with pcDNA3.1-E2 and pcDNA3.1-E2-UL49 were detected by immunofluorescent antibody test and Western-blotting. The cells containing pcDNA3.1-E2 were IFA-positive to CSFV antisera and the expressed protein E2 was found around the nucleus, but the expressed fusion protein E2-VP22 was located both in the cytoplasm and around the nucleus. Western-blotting analysis indicated that both the protein E2 and the fusion protein E2-VP22 could react with CSFV specific antibodies and the E2-VP22 could also react with BHV-1 specific antibody. The expression level of the E2-VP22 protein gene was higher than that of the E2 gene（P<0.01）. The results suggested that the VP22 protein had the function of protein transduction. |
|
|
|
Key words: classical swine fever virus; E2 gene; UL49 gene; expression in vitro |
|
|
|
|
|
《中国兽医科学》 ©
版权所有
Chinese Veterinary Science
zgsykx@zgsykx.com |
