Construction of a CD133 lentiviral expression vector and establishment of its stably transfected SGC7901 cell line

Objective To construct a CD133 lentiviral expression vector and to establish its stably transfected SGC7901 cell line.Methods Full-length CD133 gene amplified by reverse transcription-poly-merase chain reaction (RT-PCR) was inserted into a pGMLV-PB1-1 vector to construct pGMLV-PB1-1-CD133 and pGMLV-PB1-1 vectors,and then transfected into 293T cells to precede the lentivirus equipment package.Subsequently,we collected the lentivirus venom to infect the SGC7901 cells and establish a stably overexpressed cell line named SGC7901-CD133.Results A CD133 lentiviral expression vector (pGMLV-PBl-1-CD133) was successfully constructed by restrictive enzyme digestion and plasmid sequencing.RT-PCR and Western blotting revealed the increased mRNA and protein expression of CD133 gene in cells transfected with pGMLV-PB1-1-CD133 (1.160 ± 0.051,0.835 ± 0.077) as compared with the control groups.Therefore the SGC7901-CD133 cell line was successfully established through the experiment.Conclusion A lentiviral CD133 expression vector and its SGC7901 cell line were successfully constructed. Key words: Gastric adenocarcinoma;  CD133;  Lentiviral vector;  Transfection