Construction of eukaryotic expression vectorinsertedby MafA and its expression in HepG-2 cells
- VernacularTitle:MafA真核表达载体的构建及在人肝癌细胞中的表达
- Author:
Dong HUANG
- Publication Type:Journal Article
- Keywords:
pEGFP-N2;
pEGFP-C1;
Mus musculus v-maf musculoaponeuro-tic fibrosarcoma oncogene family,protein A(MafA)
- From:
Journal of Chongqing Medical University
2007;0(08):-
- CountryChina
- Language:Chinese
-
Abstract:
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.
