关键词: 囊膜糖蛋白Ｅ0基因; 克隆; 序列测定; 增强型绿色荧光蛋白

Abstract: E0 gene was taked back from the pGEM-T Easy-E0 recombinant plasmid cleaved by BamH I and Pst I, was cloned into the EGFP. High pure recombinant plasmid of pEGFP- E0 was transfected into PK-15 cell, BHK-21 cell, VERO cell by LipofectamineTM2000, examine the cell for fluorescence using a UV microscope. All could find the fluorescence in PK-15 cells, BHK-21 cells, VERO cells. In 24 h, 48 h and 72 h, count of the the cell for fluorescence were increasing, count of the cell for fluorescence were most in PK-15 cells, BHK-21 cells in 72 hs, VERO cells. count of the cell for fluorescence were most in PK-15 cells in 72 h. Detection indicated expressed E0 protein mainly locate in the plasma of the PK-15 cells. After cleaved by BamH I and Pst I, PCR and indirect immuno fluorescence assay, detection indicated large number of expressed E0 protein are in the PK-15 cells.After PK-15 cells and PK-E0 cells infected by HCLV, the titer of HCLV was the highest in 72 h, then began to decrease. Detection indicated large number of expressed E0 protein caused increase of the titer of HCLV.

Key words: glycoprotein E0; clone; sequence analysis; enhance green fluores-cent protein 