【摘要】 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NeuritincDNA的开放阅读框克隆到真核表达载体pcDNA4.0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。更多还原

【Abstract】 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.更多还原