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.
出处
《黑龙江畜牧兽医》
CAS
北大核心
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)
