Expression and purification of human apolipoprotein M / 中南大学学报(医学版)

ABSTRACT

OBJECTIVE@#To express and purify the extra cellular full-length human apolipoprotein M(ApoM).@*METHODS@#The ApoM gene fragment was amplified from the human liver cDNA library by PCR. The resulting product was cloned into pGEXT vector and sequenced. Then the confirmed canstatin cDNA was cloned into plasmid E.coli JM109 and then transformed into E.coli DL21(DE3) where it was induced to express protein by IPTG.@*RESULTS@#The ApoM gene was cloned by PCR and a 560 bp DNA fragment was shown on the agarose electrophoresis. The cloned gene was sequenced and demonstrated to have the same sequence as that of human ApoM gene in GenBank. Then ApoM cDNA gene fragment was induced by IPTG, and a 24 kD recombinant ApoM protein was tested on SDS-PAGE.@*CONCLUSION@#Human ApoM gene is successfully cloned and its recombinant proteins are expressed.