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目的:构建携带人神经生长因子(h N G F) 基因的真核细胞表达载体,为应用 N G F 进行老年性痴呆( A D) 等疾病基因治疗打基础。方法:应用基因重组技术将h N G Fc D N A 克隆到逆转录病毒载体p L X S N 中,用限制性内切酶酶切分析重组质粒p L N G F S N 中h N G F基因的插入方向,用 P C R、斑点杂交和 Southern 杂交对重组质粒p L N G F S N 作进一步鉴定。结果:h N G Fc D N A已正确地克隆到逆转录病毒载体p L X S N 中,而构建成重组逆转录病毒载体p L N G F S N。结论:真核细胞表达载体p L N G F S N 的成功构建,为进一步开展 N G F基因治疗 A D 等中枢神经系统疾病奠定了基础。
OBJECTIVE: To construct eukaryotic expression vector carrying human NGF (hNGF) gene and lay the foundation for the gene therapy of diseases such as senile dementia (AD) using NGF. METHODS: The hNG Fc D N A gene was cloned into the retroviral vector p L X S N using gene recombination technique. Restriction endonuclease digestion was used to analyze h N G F in the recombinant plasmid p L N G F S N The insertion direction of the gene was further identified by PCR, dot blot and Southern hybridization. The recombinant plasmid pL N G F S N was further identified. Results: The hNG Fc D N A was correctly cloned into the retroviral vector p L X S N and constructed into the recombinant retroviral vector p L N G F S N. CONCLUSION: The successful construction of eukaryotic expression vector pLNG F S N lays the foundation for the further development of NGF gene therapy for central nervous system diseases like AD.