Construction of adenoviral expression vector carrying FLIP gene and its expression in rat pulmonary vascular endothelial cells
AIM: To construct an adenoviral vector carrying rat FLIP (FLICE-inhibitory protein) gene and to observe its expression in rat pulmonary vascular endothelial cells (PVEC). METHODS: Recombined adenovirus Ad-FLIP was constructed by Ad-max system. The PVECs of rats were isolated, primarily cultured in vitro and infected with Ad-FLIP. The expression of FLIP was detected by Western blot and RT-PCR. RESULTS: FLIP gene was successfully cloned and incorporated into recombinant adenovirus Ad-FLIP. The expression of FLIP gene in the infected cells was detected by RT-PCR. The cells were infected with Ad-FLIP and FLIP protein was detected in the ingredients ofcell disruption. CONCLUSION: The recombinant adenoviral vector carrying rat FLIP gene is successfully constructed and Ad-FLIP efficiently elevates the expression of FLIP gene and protein in cultured rat PVECs.