HUANG Dong, et al. Construction of eukaryotic expression vectorinsertedby MafA and its expression in HepG-2 cells[J]. Journal of Chongqing Medical University, 2009,34(8):988-991.
HUANG Dong, et al. Construction of eukaryotic expression vectorinsertedby MafA and its expression in HepG-2 cells[J]. Journal of Chongqing Medical University, 2009,34(8):988-991.DOI:
Objective:To construct eukaryotic expression vectors pEGFP-N2/MafA and pEGFP-C1/MafA
encoding the mouse musculus v-maf musculoaponeurotic fibrosarcoma oncogene family
protein A(MafA)gene
for the further research of its expression in HepG-2 cells.Methods:pRNA of the mouse MafA gene was distilled by RT-PCR from the mouse
and inserted into Hind Ⅲand SalⅠrestriction sites by PCR
then cloned into pEGFP-N2 and pEGFP-C1 vectors to obtain plasmids pEGFP-N2/MafA and pEGFP-C1/MafA.Human liver cancer cells(HepG-2) were transfected with formed plasmids by means of lipidosome.The MafA-EGFP fused protein was viewed directly with fluorensce microscope
and expression of MafA and insulin Ⅱ was detected by RT-PCR.Results:The mouse MafA gene was amplified through RT-PCR and successfully cloned into transfer vectors.The favorite gene sequence could be expressed and was completely consistent with that reported in genebank
but the insulin Ⅱ gene expression was not detected.Conclusion:The recombinant plasmids pEGFP-N2/MafA and pEGFP-C1/MafA were successfully constructed.But the insulin Ⅱgene was not expressed.