构建人类neuritin真核表达系统

目的：构建人类neuritin基因真核表达载体，并观察其转染的PC12细胞中neuritin的表达。方法：用基因重组技术，将人类Neuritin cDNA的开放阅读框克隆到真核表达载体pcDN A4．0中，经酶切鉴定及测序分析、并以脂质体介导转染PC12细胞，了解其在细胞内的表达。结果：酶切鉴定及测序分析，表明重组pcDNA4．0-neuritin表达质粒克隆成功．neuritin基因在PC12细胞中获得表达。结论：以脂质体介导pcDNA4．0-neuritin质粒转染真核细胞，为基因治疗神经系统退行性病变奠定实验基础。

Human Neuritin Cloning and Its Expression in Eukaryotic Cells

Objective: To construct the eukaryotic expression vector of human neuritin and express it in PC12 cells.Methods: By gene recombination technique,the coding sequence of human neuritin was cloned into pcDNA4.0 eukaryotic expression vector.After analysis by restriction enzyme digestion and DNA sequencing,the recombinant pcDNA4.0-neuritin plasmid was transfection into PC12 cells by using catiomic lipid ( Tfx TM).Neuritin protein was then determined by using pcDNA4.0-neuritin plasmid transfected eukaryotic cells.Results: Analysis by restricting enzyme digestion and DNA sequencing of pcDNA4.0-neuritin recombinant showed that neuritin cloning was successful. The recombinant plasmid can express active neuritin protein in eukaryotic cells.Conclusion: Further study on the role of both neuritin and gene therapy is significant in the treatment of neurodegenerative diseases.