Expression of Human Globular Adiponectin-Glucagon-Like Peptide-1 Analog Fusion Protein and Its Assay of Glucose-Lowering Effect In Vivo

• Main Author: Tongfeng Zhao, Jing Lv, Jiangpei Zhao, Xiao Huang, Haijuan Xiao
• Format: Article
• Language: English
• Published: Ivyspring International Publisher 2011-01-01
• Series: International Journal of Medical Sciences
• Online Access: http://www.medsci.org/v08p0203.htm

<p>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 <i>Escherichia coli</i> BL21 (DE3). A recombinant fusion protein of about 25KD was expressed from BL21 (DE3) cells after isopropylthio-&#946;-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.</p>