title: Expression of human globular adiponectin-glucagon-like peptide-1 analog fusion protein and its assay of glucose-lowering effect in vivo.
abstract: 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.
authors: Department of Geriatrics, the Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310000, PR China. zhaotongfeng@yahoo.com.cn
authors: Tongfeng
authors: T
authors: Zhao
authors: Jing
authors: J
authors: Lv
authors: Jiangpei
authors: J
authors: Zhao
authors: Xiao
authors: X
authors: Huang
authors: Haijuan
authors: H
authors: Xiao
mesh_terms: D052242:Adiponectin / Q000008:administration & dosage / Q000235:genetics / Q000627:therapeutic use; D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D001786:Blood Glucose / Q000187:drug effects*; D003921:Diabetes Mellitus, Experimental / Q000097:blood / Q000188:drug therapy*; D004765:Enteropeptidase / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D052216:Glucagon-Like Peptide 1 / Q000008:administration & dosage / Q000031:analogs & derivatives / Q000235:genetics / Q000627:therapeutic use*; D006801:Humans; D002479:Inclusion Bodies / Q000378:metabolism; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D010446:Peptide Fragments / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology / Q000627:therapeutic use*; D014170:Transformation, Genetic
chemical_list: C499918:ADIPOQ protein, human; D052242:Adiponectin; D001786:Blood Glucose; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D052216:Glucagon-Like Peptide 1; D004765:Enteropeptidase