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

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 neurotroph ic 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GDNF DNA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。

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 nueleotide 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.