Neurturin嵌合神经营养因子重组腺病毒的构建及体外活性测定  

The Construction of Recombinant Adenovirus Delivering Chimeric Neurturin and Activity Assay in vitro

作　　者：王文举[1] 孙茂盛[1] 严敏[1] 李鸿钧[1] 

机构地区：[1]中国医学科学院/北京协和医学院医学生物学研究所,昆明650118

出　　处：《中国生物工程杂志》2010年第9期7-12,共6页China Biotechnology

基　　金：国家自然科学基金(30971003);云南省基础研究重点项目(2007C0012Z);云南省中青年学术技术带头人后备人才项目(2005PY01-24)资助项目

摘　　要：Neurturin是神经营养因子家族成员,研究发现体外表达的重组Neurturin能够促进多巴胺能神经元的存活,对帕金森氏病的基因治疗有重要意义。但有研究指出野生型Neurturin在细胞中不能完成加工进而不能被分泌到胞外。因此实验对野生型Neurturin进行改造,将NGF的信号肽与Neurturin的成熟肽进行拼接,形成一种嵌合的NGF/NTN基因,并将该基因插入重组腺病毒基因组中,在AD-293细胞中包装获得重组腺病毒。通过免疫荧光和Westernblotting证实Neurturin能够在胞内表达,并且能够对其进行加工最后分泌至胞外。最后通过鸡胚背根神经节培养实验证明该嵌合基因能够表达具有明显活性的Neurturin。Neurturin is a member of neurotrophic family,and the gene engineering product of the Neurturin can promote the survival of the dopaminergic neurons,which is a new approach to the therapy of Parkinson＇s disease.However,a few researches have indicated that the wild type of Neurturin cannot be processed and secreted into the medium.Therefore,the study intends to rebuilt the gene of wtNeurturin and replaced the signal peptide of wtNeurturin by the counterpart of NGF.The hybrid Neurturin was inserted into the genome of adenovirus and packaged in AD-293 cells.The chimeric Neurturin could be expressed in cytoplasm of vero cells and be processed and secreted,which was detected by immunofluorescence and Western blotting.Finally,the culture of dorsal root ganglia of chick embryo test had indicated that Neurturin expressed by chimeric gene had good biological activity.

关 键 词：NEURTURIN 嵌合基因 重组腺病毒 体外活性 帕金森氏病 

分 类 号：Q7[生物学—分子生物学] R74[医药卫生—神经病学与精神病学]