人源μ型阿片受体的cDNA克隆及转染CHO细胞的活性分析  

Expression of Human μ-opioid Receptor cDNA in CHO Cell

作　　者：刘威[1] 段海清[1] 李淑琴[1] 刘秀丽[1] 张兆山[1] 

机构地区：[1]北京生物工程研究所,北京100071

出　　处：《生物工程学报》2004年第3期372-376,共5页Chinese Journal of Biotechnology

摘　　要：μ型阿片受体是阿片类药物镇痛与成瘾的分子基础。从人脑组织总RNA通过RT PCR扩增获得 μ型阿片受体的cDNA ,将其克隆至pcDNA3 1(+)中 ,用酶切鉴定正确的重组质粒转染CHO细胞。筛选的单克隆细胞株 ,检测阳性的细胞克隆表达的 μ型阿片受体介导胞内信号转导的能力。通过与激动剂和拮抗剂的信号转导分析证实 ,阳性的细胞克隆表达的 μ型阿片受体与天然的 μ型阿片受体具有基本一致的生物学特性 ,因此可以用来作为高效镇痛低成瘾药物筛选平台的候选细胞株。Opioid receptor, is classified into three subtypes,μ、κ and δ,with the μ-type receptor plays important roles in opioid analgesia and opioid addiction. The cDNA encoding μ-type receptor was obtained by RT-PCR from human brain RNA and was cloned into pcDNA3.1(+). The resultant recombinant plasmid pcDNAMORs were transfected into CHO cells by liposome. After PCR identification, the positive clone were treated with agonist and antiagonist were tested for their competence of signal transduction.CHO cells that contained μ-opioid receptor in the expression vector pcDNA3.1(+) acquired naloxone-blockable high-affinity specific binding of morphine and DAMGO. The concentration of cAMP in CHO cells transfected with pcDNAMOR was reduced after binding to morphine and DAMGO, and increased after binding naloxone. These results indicate that the μ-type receptor expreesd on the CHO cell has similar biological property as the nature receptor.The availability of these specific cell lines will facilitate the drug development and promote our understanding the mechanism underlying opiate addiction.

关 键 词：Μ型阿片受体 信号转导 受体-配体结合 

分 类 号：R346[医药卫生—基础医学]