Research On The Cloning, Expression & Function Of HLIF

Objective: To clone human LIF (hLIF) gene and construct its eukaryotic expression vector, express it in COS-7cells and ECV-304 cells and explore the effect of LIF on HL-60 cells ,study the effect of the ECV-304 cells modified with LIF gene on the ex vivo culture of HSC/HPC in cord blood.Methods: The cDNA fragment encoding LIF was amplified by RT-PCR .Then the recombinant eukaryotic expression plasmids were constructed and identified by endonucleases digestion and PCR. The cDNA fragment was sequenced. The COS-7 cells and ECV-304 cells were infected by Eukaryotic Expression plasmid pcDNA3.0-LIF respectively,then the positive ECV-304 cells were obtained by selected with G418. The effect of rhLIF on HL-60 cells was detected by means of MTT FCM. The Synergistic effect of rhLIF and IL-24 gene on apoptosis of HL-60 cells was observed by LSCM FCM and immunohistochemistry stain assay. Further investigation was made to study the effect of the ECV-304 cells modified with LIF gene on the ex vivo culture of CD34+cells in cord blood.Results: The full encoding sequence of LIF was obtained and identified with that included in GenBank. Expression vectors were...