EBNA2 gene eukaryotic expression vector and expression of functional products.docVIP

 PAGE \* MERGEFORMAT 13 EBNA2 gene eukaryotic expression vector and expression of functional products [Keywords:] EBNA2 Construction of eukaryotic expression vector of EBNA2 and functional detection of its product [Abstract] AIM: To construct the eukaryotic expression vector of EBNA2 and to express it in eukaryotic cells and detect the function of its product. METHODS: EBNA2 gene was obtained by PCR and DNA recombination from the plasmid pSG5EBNA2. PCR product of EBNA2b was then inserted into plasmid pMD18T and sequenced. The full length EBNA2 gene was cut and inserted into eukaryotic expression plasmid pCMVHA. Using LipofectamineTM 2000, the plasmid pCMVEBNA2 was transfected into Cos7 and then detected by Western blot and immunofluorescence. The activity of EBNA2 was detected by luciferase reporter assay. RESULTS: The EBNA2 gene was successfully cloned and its sequence was identical to that in GenBank. After plasmid pCMVEBNA2 was transfected into Cos7, the expressed product of EBNA2 was detected by Western blot and immunofluorescence. The transcriptional activity of EBNA2 was demonstrated by reporter assay. CONCLUSION: The eukaryotic expression vector pCMVEBNA2 has been successfully constructed and successfully expressed in eukaryotic cell line. The EBNA2 protein has transcriptional activation. [Keywords] EBNA2; clone; expression; genetic vector [Abstract] Objective: To construct EB virus core antigen 2 (EBNA2) eukaryotic expression vector, expressed in eukaryotic cells, and to detect its expression product features. Methods: Recombinant DNA and the PCR method from the plasmid subclones pSG5EBNA2 in EBNA2 encoding genes, inserted into the eukaryotic expression vector pCMVHA. application of the method of liposome recombinant plasmid pCMVEBNA2 transiently transfected Cos7 cells by Western blot and immunofluorescence used to detect EBNA2 expression. application of luciferase reporter protein experiments detection of EBNA2 activity. Results: The cloned the ge