| 人GLP-1R基因转染BHK细胞及重组细胞的应用 |
| |
| 引用本文： | 朱振洪,王同映,黄岩山. 人GLP-1R基因转染BHK细胞及重组细胞的应用[J]. 生物技术通报, 2007, 0(3): 118-121 |
| |
| 作者姓名： | 朱振洪 王同映 黄岩山 |
| |
| 作者单位： | 1. 浙江中医药大学生物工程学院,杭州,310053
2. 杭州九源基因工程有限公司,杭州,310018 |
| |
| 摘 要： | 为了构建表达人胰高血糖素样肽-1受体(GLP-1R)基因的BHK细胞株,并利用该重组细胞对GLP-1等相关肽进行活性测定,首先通过酶切、连接方式将人GLP-1R基因克隆至真核表达载体pCDNA3.(1 )中,然后用脂质体转染法将重组质粒转染至BHK-21细胞,转染后的细胞经G418加压筛选、细胞有限稀释等方法获得克隆细胞株。经过该细胞株RT-PCR验证,结果证实目的基因已整合至BHK-21细胞基因组中,并获得成功转录和表达。活性检测实验表明该重组细胞株经过GLP-1的刺激后,其细胞中的cAMP含量得到明显提升。该细胞株的构建为GLP-1及相关肽的活性测定奠定了基础。
|
| 关 键 词： | GLP-1受体 |
| 修稿时间： | 2006-12-27 |
Construction of Recombinant BHK/GLP-1R Cells and its Application |
| |
| Zhu Zhenhong,Wang Tongying,Huang Yanshan. Construction of Recombinant BHK/GLP-1R Cells and its Application[J]. Biotechnology Bulletin, 2007, 0(3): 118-121 |
| |
| Authors: | Zhu Zhenhong Wang Tongying Huang Yanshan |
| |
| Affiliation: | 1.Zhejiang Chinese Medical University ,Hangzhou 310053;2Hangzhou Jiuyuan Gene-engineering Co. ,Ltd,Hangzhou 310018 |
| |
| Abstract: | In order to establish a recombinant BHK-GLP1R cell line which has a good application to exam biological activity of GLP-1 and its analogues.The human glucagons-like peptide 1 receptor(GLP-1R)gene was cloned into eukaryotic expression vector pCDNA3.1( ).The recombinant expression vector was transfected into BHK-21 cells by mean s of lipofectin procedure.After selected by G418,the positive cell clones were analysed by RT-PCR to confirm the steady expression of GLP-1R gene in BHK cells.The analysis of RT-PCR show that GLP-1R gene had been integrated into BHK cell genome and could express the GLP-1R protein successfully in BHK cells.The recombinant cells had been applied to exam the quantity of intracellular cAMP after stimulated by GLP-1 or its analogues. |
| |
| Keywords: | BHK-21 cAMP |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
|
