Biotechnology Bulletin ›› 2015, Vol. 31 ›› Issue (3): 218-222.doi: 10.13560/j.cnki.biotech.bull.1985.2015.04.032

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Preparation of Prokaryotic Expression Construct of Human FGF21 cDNA and Its Recombinant Protein Expression

Zhang Lilin1 Tang Qinglan2 Xu Qingzhong1 Lei Tingwen1 Li Hongmei1   

  1. (1. Department of Biochemistry and Molecular Biology,Guiyang Medical College,Guiyang 550004;2. Department of Clinical Laboratory,the Third People’s Hospital of Hainan Province,Sanya 572000)
  • Received:2014-09-25 Online:2015-03-16 Published:2015-03-16

Abstract: Preparation of prokaryotic expression constructs of human FGF21(fibroblast growth factor,FGF)cDNA and induction of recombinant hFGF21 protein expression. Total RNA was extracted from human liver, and the target cDNA fragment was obtained using RT-PCR. The amplified cDNA fragment was cloned into pMD19-T for preservation. Then the expression construct pET-28a(+)-hFGF21 was successfully constructed and expressed with IPTG induction, and the his-hFGF21 protein was purified with histide-selective nickel affinity gel and identified by Western blot. Western blot analysis showed that the fusion protein had specific binding with a FGF21 antibody.

Key words: human FGF21, molecular cloning, prokaryotic expression, protein purification