激光生物学报2017，Vol.26Issue(6)：534-539,6.DOI:10.3969/j.issn.1007-7146.2017.06.010
GIP受体激动剂类药物筛选模型的构建
Construction of Screening Model for GIP Receptor Agonists
摘要
Abstract
GIP receptor agonists which can stimulate insulin secretion could be potential drugs in the treatment of type II diabetes. Therefore, it is important of the screening model for GIP receptor agonists. In this study, GIPR gene was amplified by PCR, digested by HindIII/XhoI and then connected to the expression vector pCMV6-AC-GFP to construct pCMV6-AC-GIPR-GFP recombinant plasmid. The recombinant plasmid was transformed into Escherichia coli DH5α, subsequently confirmed by colony PCR, restriction endonuclease digestion and sequencing. The recombinant plasmid was transfected into RIN-m5F cells by lipofectamine 2000. After screening with G418, the monoclonal RIN-m5F/GIPR-GFP cell line was selected. The results showed that the GIPR gene with a length of 1319 bp was amplified and cloned into the eukaryotic expression vector of pCMV6-AC-GFP. After analysis of colony PCR, enzyme digestion and sequence, the construction of plasmid pCMV6-AC-GIPR-GFP was proved to be successful. Lots of positive puncta were observed in the treatment of GIPR agonist of ( D-Ala2 ) GIP . The results reveal that GIPR-GFP cell line with stable expression was successfully constructed. This cell line can be used to screen agonists of GIPR for exploring potential new medicines for diabetes.关键词
GIP受体激动剂/药物筛选模型/GFP/RIN-m5F/质粒转染Key words
GIPR agonist/drug screening model/GFP/RIN-m5F/plasmid transfection分类
生物科学引用本文复制引用
周海燕,吴艳阳,刘东波,谢红旗..GIP受体激动剂类药物筛选模型的构建[J].激光生物学报,2017,26(6):534-539,6.基金项目
国际科技合作与交流专项(2013DFG32060) （2013DFG32060）
