Construction of eukaryotic expression vector containing latent membrane protein 1 and its expression in human nasopharyngeal carcinoma cells

Objective:To construct a eukaryotic expression vector containing latent membrane protein 1(LMP1) and to detect its expression in human nasopharyngeal carcinoma CNE2 cells.Methods:The cDNA segment encoding LMP1 from human nasopharyngeal carcinoma cell line C15 was acquired and cloned into a eukaryote expression vector pEGFP-C1 after double digestion.The recombinant vector was identified by restriction enzyme analysis and nucleotide sequence determination.The eukaryotic expression plasmid pEGFP-C1-LMP1 was transiently transfected into CNE2 cells.The location and expression of LMP1 was detected by laser scanning confocal microscope.Results:Digested with SalⅠ and BglⅡ,the recombinant vector was examined by agarose gel electrophoresis,and the result was in accordance with the anticipated objective strap size.The LMP1 was correctly inserted into pEGFP-C1;the sequencing results were identical with those reported in GenBank.GFP-LMP1 could be detected in both cell membrane and cytoplasm of CNE2.Conclusions:Recombinant plasmid pEGFP-C1-LMP1 is constructed successfully in vitro,which can also express in the cytoplasm of CNE2 in human nasopharyngeal carcinoma.