Fudan University Journal of Medical Sciences 2010;37(1):76-79,99

Construction of inducible lentiviral expression vector containing MST1 gene and its expression in Huh-7 cell line

Bo GU

Keywords

inducible lentiviral vector; MST1 gene; doxcycline

Country

China

Language

Chinese

Abstract

Objective To construct inducible lentiviral vector containing MST1 gene, which provides an ideal model for further study of the relationship between liver cancer and MST1 gene. Methods We cloned MST1 into inducible lentiviral vector. Two lentiviral vectors (pLVPT-tTRKRAB-MST1 and pLV-tTRKRAB-Red) with package plasmids were contransfected into 293FT respectively, and the lentiviral viruses were harvested from 293FT. Viruses were used to infect liver cancer cell line (Huh-7) in tandem. We used doxcycline to induce the expression of target gene MST1 which was indentified by Western blot after 7-day cell culture. Results The recombinant inducable lentiviral vector containing MST1 gene was successfully constructed. The lentiviruses were also obtained and mediated by 293FT, which were highly efficient to infect liver cell lines Huh-7. The expression of MST1 was identified under Dox induction. Conclusions The recombinant inducable lentiviral vector containing MST1 gene has been successfully constructed. It is viable to obtain inducible cell lines Huh-7 with MST1 gene expression under Dox induction.