野鸭源新城疫病毒V蛋白基因在DF-1细胞中的表达及V蛋白的亚细胞定位

Expression of V protein gene of mallard-origin Newcastle disease virus in DF-1 cells and sub-cellular localization of the expressed V protein

为研究野鸭新城疫病毒V蛋白基因在真核细胞中的表达及V蛋白的亚细胞定位,将新城疫病毒V基因片段与pcDNA3.1载体连接,构建pcDNA3.1-V重组表达质粒,经脂质体转染DF-1细胞后,用Western-blot验证该基因在DF-1细胞中的表达;用激光共聚焦显微成像技术确定V蛋白在真核细胞中的分布及亚细胞定位。结果显示,重组质粒pcDNA3.1-V在外源真核细胞DF-1中获得了表达,表达蛋白主要聚集于细胞质。研究结果为进行野鸭源新城疫病毒V蛋白的功能研究奠定了基础。

The aim of the study was to study the expression and sub-cellular localization of V protein gene of Newcastle disease virus in eukaryotic cells.The recombinant plasmid pcDNA3.1-V was constructed and transfected into DF-1cells with Lipofectamine 2000.Western-blot was used to verify the expression of V protein gene in DF-1cells.The distribution and sub-cellular localization of the expressed V protein in DF-1cells was determined using laser confocal microscopy imaging technique.In result,the recombinant plasmid was successfully expressed in DF-1cells and the V protein was mainly distributed in the cytoplasm.The result laid the foundation for studies on functions of V protein of mallard-origin Newcastle disease virus.