Construction of pcDNA3.1-3a in pcDNA3.1 expression vector and its expression in neuroblastoma cell line SK-N-SH cells

Objectives To clone and express the pcDNA3.1-3a gene fused with myc-6his tag in pcDNA3.1 eukaryotic expression system and to identify the protein of fused expression in order to derive the fused gene which may express the methyltransferase catalysis domain.Methods The methyltransferase catalysis domain 3a fused with myc-6his was derived from pcDNA4.0-GBD-3a-myc-6his plasmid with PCR amplification,and cloned into the eukaryotic expression vector pcDNA3.1.The vector was identified by PCR and double enzymes digestion.The expression product was analyzed with Western blot in Neuroblastoma cell line SK-N-SH cells.Results The pcDNA3.1-3a gene was amplified with PCR,and the interest protein could be specifically recognized by anti-myc-tag monoclonal antibody in the SK-N-SH cells.Conclusion The vector pcDNA3.1-3a may be cloned into the eukaryotic expression vector correctly and expressed successfully in Neuroblastoma cell line SK-N-SH cells.