Construction and Expression of Recombinant Wild Lipoprotein Lipase Gene Plasmid / 上海第二医科大学学报

ABSTRACT

Objective To study the construction of recombinant wild lipoprotein lipase(LPL) gene plasmid and its expression in COS-1 cells. Methods The LPL cDNA was isolated from the human epiploon adipose tissue by means of RT-PCR.The LPL cDNA was ligated into the pcDNA3.1Zeo(+).The recombinant pcDNA3.1Zeo(+)-LPL cDNA was identified by endonucleases,PCR and DNA sequencing.COS-1 cells were transfected with the recombinant LPL gene plasmid using Lipofectamine 2000~(TM).The LPL mass in cells and the culture medium were determined by a Markit-M LPL Kit.Spectrophotometry was used to measure the LPL activity. Results The LPL gene was ligated into the pcDNA3.1Zeo(+) plasmid identified by endonucleases and PCR.The sequence of the LPL gene was the same as the sequence of the Gene Bank identified by DNA sequencing.Wild pcDNA3.1Zeo(+)-LPL(cDNA) plasmids was transformed into the COS-1 cells. Conclusion The recombinant plasmid pcDNA3.1Zeo(+)-LPL cDNA could be constructed and successfully transformed into the COS-1 cells.