μ型阿片受体的克隆及序列分析

Cloning and sequencing of μ opioid receptor

目的 为实现人μ型阿片受体在哺乳动物细胞中的表达以及研究其生物学特性、下游基因表达及功能研究,克隆了人全长μ型受体cDNA。方法 从人脑组织中提取总RNA ,通过逆转录巢式PCR ,扩增μ受体全长cDNA。克隆至pMD1 8 -T载体中,并进行了酶切、PCR鉴定及测序分析。结果 获得大小1 2 2 9bp大小的μ受体全长cDNA基因,并构建到载体中,为研究受体信号转导下游基因的表达奠定理论基础。结论 利用巢式RT PCR法成功克隆了的μ受体全长cDNA序列,构建了pT -MD/hμR质粒。

Objective : To study the biological activities, expression and function of downstream related genes in mammary culture cells, human μ opioid receptor cDNA is cloned. Methods: Human μ opioid receptor cDNA was amplified from fetal brain total RNA by reverse transcriptional nest-PCR. The inserted cDNA into pMD18-T vector was identified by endonuclease digestion,PCR and sequencing. Results: The positive recombinant cloning of μ opioid receptor cDNA in 1229bp laid the foundation of further research on expression and activity of human μ receptor. Conclusion: μ opioid receptor cDNA is cloned by nest RT-PCR and the recombined plasmid pT-MD/hμR is constructed successfully.