Construction of Eukaryotic Expression Vector Containing the HHV—8 K 12 and Green Fluoresent Protein Gene

Objective To construct eukaryotic expression vector containing the HHV-8 K12 gene and green fluorescent protein gene.Methods The K12 gene of HHV-8 digested with EcoRI and XhoI was inserted into MCS sites of eukaryotic expression vec-tor pCR3.1 to construct pCR-K12;Green fluorescent protein(GFP) gene was amplified by polymerase chain reaction(PCR) from pGF-Puv as template, digested with KpnI and EcoRI and then inserted into upstream of K12 of pCR-K12 to construct pCR-GFP-K12 .Results Restriction endonucleases digestion of pCR-GFP-K12 was the same as expected, in which K12 was fused with GFP and con-trolled by promoter CMV.Conclusion Construction of pCR-GFP-K12 was successful ,which would benefit study on biology active-ty and action mechanism of HHV-8 K12.