神经损伤与功能重建Issue(5)：361-364,4.DOI:10.3870/sjsscj.2014.05.001
人神经生长因子重组慢病毒载体构建及检测
Construction and Identification of Lentiviral Vector Carrying Nerve Growth Factor Gene
摘要
Abstract
Objective:To construct the recombinant lentiviral vector containing nerve growth factor (NGF) gene and test the overexpression of NGF gene in 293T cells. Methods:The NGF gene was cloned into lentiviral shuttle plasmid(Ubi-MCS-3FLAG and-SV40-EGFP), and identified by the enzyme cut assay, PCR and gene sequencing. It was then transfected into 293T cells to construct the recombinant lentiviral vector plasmid. The infection efficiency of lentivrius transfection was detected by fluorescence microscope and the protein expression of NGF was tested by Western blot. Results:The construction of lentiviral vector carrying NGF gene was correct which was verified by the enzyme cut assay, PCR and gene sequencing. Strong fluorescence was observed in 293T cells after lentiviral vectors transfection. Western Blot analysis showed a characteristic band of NGF.Conclusion:Recombinant lentiviral vector carrying NGF gene was constructed successfully.关键词
神经生长因子/慢病毒载体/糖尿病神经源性膀胱Key words
nerve growth factor/lentiviral vector/diabetic neurogenic bladder分类
医药卫生引用本文复制引用
张飞,吴建红,杜义恒,刘海涛,夏术阶..人神经生长因子重组慢病毒载体构建及检测[J].神经损伤与功能重建,2014,(5):361-364,4.基金项目
国家自然科学基金 （）
