摘要
提取大鼠脑组织总RNA,通过逆转录巢式PCR,扩增μ型阿片受体全长cDNA,克隆至pMD20-T载体中,测序鉴定,纠正点突变后,经酶切连接克隆入pIRES2-EGFP中,测序及酶切结果表明μ基因正确,μ-pIRES2-EGFP质粒构建成功.用脂质体法将μ-pIRES2-EGFP转染入HEK293细胞中,在荧光显微镜下,转染细胞可以观察到绿色荧光,应用免疫组化荧光可以观察到μ基因的高强度表达.
Full length cDNA of rat μ opioid receptor was amplified from the rat brain tissue through reverse transcription and nested PCR,and cloned into pMD20-T vector. The site mutation of μ gene in the pMD20- T vector was corrected,then the μ cDNA was inserted into pIRES2-EGFP to form μ-pIRES2-EGFP recombinant eukaryotic plasmid. The μ-pIRES2-EGFP plasmid was transfected into HEK293 using Lipofectamine 2000. High expression of EGFP and μ gene was detected in HEK293.
出处
《生命科学研究》
CAS
CSCD
2009年第6期501-504,共4页
Life Science Research
基金
国家自然科学基金海外青年学者合作研究基金资助项目(30628018)
