人Parkin基因重组质粒的构建

Construction of human Parkin gene plasmid

目的应用分子生物学技术构建人Parkin基因重组质粒。方法从人类胎儿黑质组织中提取总RNA,以RT-PCR法扩增Parkin的cDNA片段;将此片段克隆于pUCm-T载体中,经过特异性限制性内切酶酶切分析片段正确后,进行全序列分析。结果克隆的cDNA与Gene Bank上的序列相比完全一致,得到了编码正确的人Parkin的cDNA全序列。结论成功构建了人Parkin基因重组质粒。

Objective To constuct human Parkin gene plasmid with molecular biological technique. Method Total RNA was extracted from human fetal Substantia nigra tissue, Parkin cDNA fragment was amplified by RT-PCR, this fragment was cloned in pGEM-T vector. The fragments was analysed after cutting by specific restriction enzyme. Result The cloned cDNA sequence was the right people Parkin coding sequence of the cDNA. Conclusion Human Parkin gene plasmid is constructed successfully.