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Int J Med Sci 2011; 8(3):203-209. doi:10.7150/ijms.8.203
Expression of Human Globular Adiponectin-Glucagon-Like Peptide-1 Analog Fusion Protein and Its Assay of Glucose-Lowering Effect In Vivo
1. Department of Geriatrics, the Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310000, PR China
In this study, human globular adiponectin-glucagon-like peptide-1 analog (gAd-GLP-1-A) fusion protein was expressed and its glucose-lowering effect was measured in vivo. We constructed a prokaryotic expression vector PET28a-gAd-GLP-1-A and transformed the vector into Escherichia coli BL21 (DE3). A recombinant fusion protein of about 25KD was expressed from BL21 (DE3) cells after isopropylthio-β-D-galactoside induction. This protein was N-terminal His-tagged gAd-GLP-1-A fusion protein. Most of the protein was expressed in inclusion body. The fusion protein in inclusion body was purified by using High-Affinity Nickel Iminodiacetic Acid Resin and refolded in urea gradient refolding buffer. The refolded protein was incubated with enterokinase to remove the N-terminal His-tag. The fusion protein without His-tag is gAd-GLP-1-A fusion protein, which exhibited significant glucose-lowering effect in diabetic mice.
Keywords: Escherichia coli, Expression, Globular adiponectin, Globular adiponectin-glucagon-like peptide-1 analog fusion protein, Glucagon-like peptide-1 analog
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How to cite this article:
Zhao T, Lv J, Zhao J, Huang X, Xiao H. Expression of Human Globular Adiponectin-Glucagon-Like Peptide-1 Analog Fusion Protein and Its Assay of Glucose-Lowering Effect In Vivo. Int J Med Sci 2011; 8(3):203-209. doi:10.7150/ijms.8.203. Available from http://www.medsci.org/v08p0203.htm