title: [Cloning, expressing of exendin-4 analogue and bioactivity analysis in vivo].
abstract: To construct, express and purify Exendin-4 analogue and detect its biological activity in vivo. Insert gene sequence into fusion partner ofpED plasmid which is helped to purification, entitled the new recombinant plasmid 5 Exendin-4 analogue polypeptide gene and fusion partner gene was linked by acid hydrolysisgene, transformed to E. coli BL21 and the fusion protein was induced by lactose. After acid hydrolysis, the Exendin-4 analogue polypeptide separated from fusion chaperon. Anion charge chromatography were used to further purification. 6 to 8 week-old ICR mice were injected (s.c) with Exendin-4 analogue, blood glucose and plasma insulin level was detected in different period after oral glucose tolerance test. The results show that high expression of inclusion body was induced by lactose, which accounted for 40% of germ proteins, the Exendin-4 analogue was obtained with the purity of 91.8% after being purified by anion charge chromatography. Bioactivity assay showed that the level of blood glucose of mouse which treated with exendin-4 analogue was obviously decreased to normal (P < 0.01), and the level of plasma insulin was increased obviously (P < 0.01).
authors.affiliation: State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 210009, Jiangsu, China. taimingli@163.com
authors.forename: Taiming
authors.initials: T
authors.lastname: Li
authors.forename: Chunjiao
authors.initials: C
authors.lastname: Gu
authors.forename: Xiaoyu
authors.initials: X
authors.lastname: Ge
authors.forename: Zhezhe
authors.initials: Z
authors.lastname: Li
authors.forename: Dan
authors.initials: D
authors.lastname: Wang
authors.forename: Yanhong
authors.initials: Y
authors.lastname: Ma
authors.forename: Tao
authors.initials: T
authors.lastname: Liu
authors.forename: Meiyou
authors.initials: M
authors.lastname: Zhang
authors.forename: Li
authors.initials: L
authors.lastname: Li
authors.forename: Jingjing
authors.initials: J
authors.lastname: Liu
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D000077270:Exenatide; D018014:Gene Transfer Techniques; D007004:Hypoglycemic Agents / Q000378:metabolism / Q000494:pharmacology; D007328:Insulin / Q000097:blood; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D010455:Peptides / Q000235:genetics / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D014688:Venoms / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology
chemical_list: D007004:Hypoglycemic Agents; D007328:Insulin; D010455:Peptides; D011993:Recombinant Fusion Proteins; D014688:Venoms; D000077270:Exenatide