The expression of E gene region of dengue 2 virus in NIH3T3 and its identification

Objective Construction of an eukaryotic expression plasmid of E gene fragment from Dengue 2 virus and eukaryotic expression of E glycosylated protein. Methods The E gene fragment of Dengue 2 virus NGC strain was amplified by RT-PCR and this fragment was cloned into eukaryotic expression vector pcDNA3. The recombinant plasmid pcDNA3-E was thus constructed and identified by sequencing and digestion with restriction enzymes. The recombinant plasmid pcDNA3-E was transfected into NIH3T3 cell by lipofectin. The expressed protein was analyzed by immunofluorescence, SDS-PAGE and Western blotting assay. Results A recombinant eukaryotic expression plasmid pcDNA3-E was successfully constructed. The recombinant plasmid expressed a 60 × 10³ protein of Dengue 2 virus. Conclusions The constructions of eukaryotic expression of E gene fragment from Dengue 2 virus and eukaryotic expression of E glycosylated protein will be helpful to study the structure and functions of the E protein, and to the development of Dengue diagnostic agents as well as Dengue virus nucleic acid vaccine.