The Experimental Study On Construction Recombinant Adenovirus Containing ApM1 Gene And Transfection Of HUVEC With It

CHAPTER ONE Cloning and eukaryotic expression of human apM1 geneObjective:To clone human apM1 gene and construct an eukaryotic expression vector of apM1 and globular adiponectin(gAdiponectin) gene, and to investigate the transfection and expression of eukaryotic expression plasmid pcdef-apM1 in HEK293 cells and pcDEF/PPT-gAdiponectin-myc -His(-)A in HUVEC cells.METHODS: RT-PCR was used to isolate, from the total RNA of human adipose tissue, the cDNA fragment encoding adiponectin, which was linked into pGEM-T Vector, then amplified, and cloned. The sequence was analyzed to identify the recombinant plasmids. Eukaryotic expression plasmid pcdef-apM1 and pcDEF/PPT-gAdiponectin-myc-His(-)A were constructed by DNA recombination. Expression vector pcdef-apM1 was transfected into HEK293 cells with Effectene reagent, and pcDEF/PPT-gAdiponectin-myc-His(-)A was transfected into HUVEC cells...