BmNPV多角体对稳定转化细胞表达荧光蛋白的包埋现象初探

为了探索家蚕核型多角体病毒(BmNPV)多角体对外源蛋白的包埋特性,构建了含有绿色荧光蛋白基因(gfp)和红色荧光蛋白基因(DsRed)的表达载体pIZT-DsRed,用该载体转染家蚕卵巢细胞系(BmN),并以吉欧霉素(zeocin)筛选得到稳定表达绿色荧光蛋白和红色荧光蛋白的BmN细胞系。进一步以野生型BmNPV感染pIZT-DsRed转化BmN细胞,发现部分多角体可以发出绿色荧光和红色荧光,Western blot检测证明多角体中含有GFP蛋白,显示BmNPV多角体可以包埋进病毒粒子以外的其他蛋白成分,表明BmNPV多角体蛋白的包埋机制中存在非特异性识别包埋的现象。

A Primary Study on BmNPV Polyhedron Embedding Fluorescent Proteins Expressed in Stably Transformed BmN Cells

To explore the embedment process of Bombyx mori nucleopolyhedrovirus (BmNPV) polyhedron,we firstly constructed an expression vector pIZT-DsRed containing both gfp and DsRed genes.This vector was used to transfect BmN cells to generate a stably transformed BmN cell line expressing both green and red fluorescent proteins through screening with zeocin.Then,the wild type BmNPV was used to infect the pIZT-DsRed transfected cell line.It was found that some of the newly formed polyhedra emitted both green and red fluorescent lights.Western blot confirmed that the polyhedra contained GFP proteins,indicating that BmNPV polyhedron proteins could embed proteins other than viral proteins into the polyhedron.This implies that a nonspecific recognition mechanism exists in the embedment of BmNPV poly-hedra.