Construction and expression of maspin/pEFIRES-N expression vector / 第三军医大学学报

ABSTRACT

Objective To construct the expression vector maspin/pEFIRES-N so as to study maspin gene on the inhibition of the growth,invasion,and metastasis in hepatocellular carcinoma.Methods The full lenth of maspin gene was cloned and ligated to the expression vector pEFIRES-N digested by EcoRⅠ+XbaⅠ by T4 DNA ligase.The recombinant vector maspin/pEFIRES-N was stably transfected into hepatocellular carcinoma MHCC-97 cell line,and the expressive changes of maspin gene were detected.Results The recombinant plasmid was amplified in the E.coli.JM109.After the identification and sequencing,the reconstructive plasmid was confirmed containing the correct and full nucleotide sequence of maspin gene and the mRNA and protein level of maspin gene were up-regulated in hepatocellular carcinoma MHCC-97 cells.The proliferation and invasion of MHCC-97 cells was inhibited.Conclusion The expression vector maspin/pEFIRES-N was constructed successfully and could be expressed in eukaryotic cells.