摘要
该文构建了一种胰高血糖素样肽-1(glucagon-like peptide-1,GLP-1)类似物活性检测的细胞模型,为GLP-1类似物的药物筛选提供了一种简单可靠的评价方法。真核表达质粒pc DNA3.1/h GLP-1R转染至中国仓鼠卵巢细胞(Chinese hamster ovary,CHO),经单克隆筛选和遗传霉素(G418)压力筛选最终筛选出6株单克隆菌株。流式细胞仪检测GLP-1受体(glucagon-like peptide-1 receptor,GLP-1R)的表达量,最终筛选获得1株高表达的细胞模型(CHO/pc DNA3.1/h GLP-1R)。RT-PCR结果显示,该细胞模型转录h GLP-1R基因;流式细胞仪检测结果和激光共聚焦结果显示,细胞模型的膜表面有h GLP-1R蛋白的表达。连续传代10次,细胞膜表面的h GLP-1R蛋白表达没有变化。构建的细胞模型活性检测结果显示,该细胞模型可以很好地用于GLP-1类似物的活性检测。综上所述,该细胞模型为GLP-1类似物的活性检测建立了一种方便可靠的体外活性检测方法。
The in vitro activity of glucagon-like peptide-1（GLP-1） analogue was detected in cell which stimulated the glucagon-like peptide-1 receptor（GLP-1R）. We constructed the cell model in Chinese hamster ovary（CHO） cells which could express h GLP-1R. Plasmid of pc DNA3.1/h GLP-1R was constructed, and then was transformed into CHO by electricity. Recombination CHO cell was selected, and confirmed by RT-PCR and flow cytometry. Results suggested that the cell model could not only transcript the gene of GLP-1R but also express the GLP-1R protein in the cell membrane. The cell model could well detect the bioactivity of GLP-1 analogue in vitro. We constructed a recombination CHO cell which could well be used to detect the bioactivity of GLP-1 analogue in vitro.
出处
《中国细胞生物学学报》
CAS
CSCD
2015年第8期1095-1101,共7页
Chinese Journal of Cell Biology
基金
国家高技术研究发展计划(863计划)(批准号:2014AA021003)
中国科学院战略性先导科技专项(批准号:XDA01040202)资助的课题~~
