人胚胎脑多巴胺D_2受体基因的克隆

Molecular Cloning of cDNA of Dopamine D_2 Receptor from Human Fetal Brain

为了克隆人多巴胺D2 受体 (D2 R)基因以用于帕金森病的治疗 ,从人胎脑中提取总RNA ,并逆转录成cDNA ;设计一对引物用PCR方法扩增目的cDNA片段 ,并将其重组于 pGEM T载体中 ,酶切鉴定插入片段后 ,进行全序列测定。结果表明从人胎脑纹状体扩增出至少 2种长度的cDNA片段 ,其中一个片段全长 1 4 4 5bp ,序列与GenBank(NM0 0 0 795 )登录的序列完全相同 ;另一片段为 1 30 5bp ,在第 6外显子开始处有 1 4 0bp的缺失 ,与已知的D2 R 3种转录体均不相同 ,可能为一种新的D2 R转录体。

In order to clone the human gene of dopamine D 2 receptor for treatment of Parkinson disease, total cellular RNA was isolated from human fetal brain and reversely transcribed into cDNA, which was amplified by PCR with the specific primers. The PCR products were inserted into pGEM-T vector for sequencing after emzymatic analysis. The results showed that two fragments were obtained. One, 1 445 bp, was identical to that in the GenBank(NM000795) and the other, 1 305 bp with a deletion of 140 bp, was different from all the three transcripts of D 2 receptors previously described and it may be a novel transcript.