2007•Zhongguo tangniaobing zazhiRequires access
Cloning of apM1 and gAdiponectin genes and construction and expression of eukaryotic expression plasmid of gAdiponectin gene
Deng Hua-cong
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Abstract
Objective To construct eukaryotic expression plasmid of gAdiponectin gene in order to study the function of adiponectin and gAdiponectin in diabetes and atherosclerosis.Methods Human adiponectin gene was cloned,then eukaryotic expression plasmid of gAdiponectin gene was constructed and transfected in HUVEC cells.Results Sequence analysis verified that the fragment cloned in pGEM-T vector was apM1 cDNA and that the fragment in pcDEF/PPT-myc-His(-)A was gAdiponectin gene.In the supernatant of HUVEC cells transfected by the plasmid,22KDa protein was detected by Western blot.Conclusions Eukaryotic expression plasmid of gAdiponectin gene is successfully constructed and expressed effectively in HUVEC cells.