1型和4型马疱疹病毒VHS蛋白在HeLa细胞中的表达与定位  

Expression and localization of the VHS proteins of equid herpesvirus type 1 and type 4 in HeLa cells

作　　者：曲娟娟[1] 胡晓亮[1] 顾海东[1] 张春苗[1] 冉多良[2] 相文华[3] 

机构地区：[1]东北农业大学生命科学学院,哈尔滨150030 [2]新疆农业大学动物科技学院,乌鲁木齐830000 [3]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,哈尔滨150001

出　　处：《黑龙江畜牧兽医》2016年第1期145-147,261,共4页Heilongjiang Animal Science And veterinary Medicine

基　　金："十二五"农村领域国家科技计划项目(2012BAD46B01)

摘　　要：为了研究1型和4型马疱疹病毒(EHV)VHS蛋白在真核细胞中的表达与亚细胞定位,试验将1型和4型马疱疹病毒VHS基因片段与p3×Flag载体连接,构建p3×Flag-VHS-1和p3×Flag-VHS-4重组表达质粒,经脂质体转染HeLa细胞后,用Western-blot验证该蛋白在HeLa细胞中的表达情况,并以激光共聚焦显微成像技术确定VHS蛋白在真核细胞中的分布及亚细胞定位。结果表明:重组质粒p3×Flag-VHS-1和p3×Flag-VHS-4在HeLa细胞中获得了表达,且主要聚集于细胞浆。说明VHS蛋白能够在外源真核细胞HeLa中表达,蛋白主要位于HeLa的细胞浆中。To study the expression and sub - cellular localization of the VHS proteins of equid herpesvirus type 1 and type 4 in eukaryotic cells, the VHS gene fragments of equid herpesvirus type 1 and type 4 was connected with p3 ×Flag vector to construct p3 ×Flag - VHS - 1 and p3× Flag- VHS- 4 recombinant expression plasmids. The recombinant expression plasmids were transfected into HeLa cells with lipofectamine 2000. Western - blot was used to verify the expression of the VHS proteins in HeLa ceils, and the distribution and sub - cellular localization of the VHS proteins in HeLa cells were determined using laser confocal microscopy imaging technique. The results showed that the p3 × Flag - VHS - 1 and p3 × Flag - VHS - 4 recombinant plasmids were successfully expressed in HeLa cells and mainly aggregated in the cytoplasm. The results indicate that the VHS proteins can be expressed in exogenous eukaryotic HeLa cells and mainly locate in the cytoplasm of HeLa cells.

关 键 词：马疱疹病毒 HELA细胞 VHS蛋白 表达 定位 免疫印迹 

分 类 号：S852.65[农业科学—基础兽医学] Q786[农业科学—兽医学]