中文摘要：

为检测a-突触核蛋白（a-Synuclein）在人神经元母细胞瘤细胞（SH-SY5Y）中的表达,构建人野生型（WT）和A53T突变型a-Synuclein质粒（p EGFP-SNCA-WT和p EGFP-SNCA-A53T）,经酶切鉴定无误后转染SH-SY5Y细胞,并对转染完成的细胞进行嘌呤霉素筛选,然后通过PCR法扩增转染后SH-SY5Y细胞的a-Synuclein片段,并对其进行测序,检测目的基因,运用Western blot法检测转染后SH-SY5Y细胞中a-Synuclein的表达。酶切鉴定结果表明p EGFP-SNCA-WT和p EGFP-SNCA-A53T质粒构建成功,嘌呤霉素筛选和转基因细胞a-Synuclein片段测序结果显示慢病毒表达载体能成功的整合到SH-SY5Y细胞基因组中,Western blot结果表明,转染后的SH-SY5Y细胞能成功的过表达a-Synuclein。a-Synuclein慢病毒表达载体构建成功,并且SH-SY5Y细胞作为宿主细胞能够表达a-Synuclein。为今后帕金森病（PD）体外模型的建立及帕金森病发病机制的研究奠定了基础。

英文摘要：

To detect the expression of α- Synuclein in SH- SY5 Y cells,the Wild- Type（ WT） or A53 T mutant α- Synuclein lentiviral expression vector（ p EGFP- SNCA- WT and p EGFP- SNCA- A53T） was constructed,the orientation of which was confirmed by restriction analysis and was transfected into SHSY5 Y cells,respectively. The individual stably transfected colony was subsequently selected in the presence of puromycin,and the gene from transfected SHSY5 Y cells was PCR amplified and confirmed by restriction analysis and DNA sequencing. Restriction endonuclease results showed that p EGFP- SNCA- WT and p EGFP- SNCA- A53 T plasmids were constructed successfully. The results from antibiotic selection and DNA sequencing showed that the lentiviral vector can be integrated into the genome of SH- SY5 Y cell successfully. Western blot assay showed that a- Synuclein could be overexpressed in the transfected SHSY5 Y cells successfully. Wild- Type or A53 T mutant a- Synuclein lentiviral expression vector was constructed successfully and SH- SY5 Y can express a- Synuclein as host cells. This paper lay the foundation for the establishment of Parkinson＇s disease（ PD） model in vitro and the research on the pathogenesis of PD.