ObjectiveTo construct herpes simplex virus type 2（HSV2） eukaryotic expression plasmid pCDNA3.0ICP27 and to evaluate its expression in Vero cells．MethodsThe target sequence of ICP27 gene was obtained and highfidelity amplified from HSV2 DNA. The ICP27 gene was cloned into a eukaryote plasmid pCDNA3.0 after restrictive endonucleases digestion．The pCDNA3.0ICP27 was checked and verified by double digestion and DNA sequence analysis. Vero cells were transiently transfected with pCDNA3.0ICP27 by lipofectamine 2 000 in vitro. RTPCR and Western blot analysis were employed to detect the expression of ICP27. ResultsThe 1741bp DNA fragment was obtained by DNA and cloned into pCDNA3.0. The recombinant plasmid pCDNA3.0ICP27 was subjected to sequence analysis which indicated all nucleotides were identical to the HSV2 ICP27 sequence provided by Genbank. Being transfected by lipofectamine 2 000, the expression of ICP27 in Vero cells was detected.ConclusionRecombinant plasmid pCDNA3.0ICP27 was constructed and expressed successfully in Vero cells．