胶质细胞源性神经营养因子基因真核荧光表达载体的构建

Construction of glial cell line-derived neurotrophic growth factor gene fluorescent eukaryotic cell expression vector

摘要: 目的: 构建可在真核细胞中表达胶质细胞源性神经营养因子(glial cell line-derived neurotrophic growth factor,GDNF)的荧光表达载体。方法: 采用聚合酶链反应(PCR)扩增GDNF基因,将GDNFDNA克隆到pEGFP-N₁质粒,通过抗性基因筛选阳性克隆,经酶切和测序鉴定。结果: 酶切和DNA序列鉴定均证实插入片段与Genebank报道的GDNF基因序列一致。结论: 成功构建了GDNF荧光真核表达载体,为从分子水平开展GDNF转基因相关研究奠定了基础。

Abstract: Objective: To construct glial cell line-derived neurotrophic growth factor（GDNF） gene fluorescent eukaryotic expression plasmid.Methods: The coding sequence of GDNF was amplified by PCR,the GDNF gene was cloned into plasmid pEGFP-N₁,and the recombinant vector was selected and identified by restriction enzyme analysis and nucleotide sequence determination.Results: Correct construction of pEGFP-N₁-GDNF was identified by methods of restriction enzyme analysis,and nucleotide sequence determination.Conclusions: An eukaryotic expression plasmid pEGFP-N₁-GDNF has been constructed successfully,which will provide the basis for studying the gene of GDNF.