IRES序列连接的人GDNF基因和EGFP基因逆转录病毒真核表达载体的构建及鉴定  

Construction and identification of recombinant vector pLXSN-EGFP-IRES2-GDNF

作　　者：陈睿 孙志军 孟宪国 刘雪平[3] 

机构地区：[1]山东医学高等专科学校康复系,山东济南250002 [2]山东医学高等专科学校医学基础部,山东济南250002 [3]山东大学山东省立医院老年神经科,山东济南250021

出　　处：《中国现代医学杂志》2010年第21期3217-3220,3225,共5页China Journal of Modern Medicine

基　　金：2006年山东省教育厅第三批科技计划项目(No:J06L85);山东省科技公关课题(No:2004GG3202013)

摘　　要：目的构建并鉴定携带人GDNF基因的逆转录病毒表达载体。方法用PCR技术从人脑胶质细胞瘤组织中扩增出GDNF全长基因,定向克隆入逆转录病毒载体pLXSN中,酶切鉴定。结果与Gene bank对照,获得正确的人GDNF序列;经酶切鉴定证实,已成功构建重组质粒pLXSN-EGFP-IRES2-GDNF。结论构建人GDNF基因的逆转录病毒表达载体可行,为下一步神经系统疾病基因治疗的研究奠定基础。【Objective】 To construct and confirm a recombinant retroviral vector carrying human GDNF gene.【Methods】 GDNF gene was amplified by PCR from glioma of human brain tissues.The gene was cloned into a retroviral vector pLXSN and recombinant vector plasmid DNA was analyzed after digested by appropriate restriction enzymes.【Results】 The correct sequence of human GDNF was obtained after compared with the sequence in GenBank.The recombinant vector pLXSN-EGFP-IRES2-GDNF was successfully constructed after confirmed by enzyme digestion analysis.【Conclusions】 Construction of eukaryotic cell expression retroviral vectors carrying human GDNF gene is feasible.It lays a foundation for further research in gene therapy of nervous system disease.

关 键 词：胶质细胞源性神经营养因子 逆转录病毒 加强型绿色荧光蛋白 

分 类 号：Q78[生物学—分子生物学]