A30Pα-synuclein原核表达载体的构建及表达

Construction of prokaryotic expression vector of A30Pα-synuclein and expression in E.coli BL21

目的构建A30Pα-synuclein基因的原核表达载体,分析其在大肠杆菌中的表达。方法酶切质粒pcDNA3.0-A30Pα-synuclein,获得A30Pα-synuclein基因的目的片段,克隆至原核表达载体pGEX-6P-1中,转化大肠杆菌BL21。IPTG诱导后,经考马斯亮蓝染色和Western-blot分析目的蛋白的表达。结果 A30Pα-synuclein基因克隆至pGEX-6P-1载体中,考马斯亮蓝染色及Western-blot检测到A30Pα-synuclein蛋白在BL21中的表达。结论成功构建A30Pα-synuclein的原核表达载体,并在大肠杆菌中表达了A30Pα-synuclein融合蛋白,为进一步研究A30Pα-synuclein在帕金森病中的作用奠定了良好基础。

Objective To construct a prokaryotic expression vector of A30P α-synuclein and analyze its expression in E.coli BL21.Methods The fragment of A30Pα-synuclein was cloned into pGEX-6P-1 vector correctly and then transformed into E.coli BL21.The expression of A30Pα-synuclein was induced with IPTG,and analyzed by Coomassie brilliant blue dyeing and Western-blot.Results A30P α-synuclein was cloned into pGEX-6P-1 properly.The recombinant fusion protein was expressed in E.coli BL21 by Coomassie brilliant blue dying and Western-blot.Conclusion The prokaryotic expression vector of A30P α-synuclein has been successfully constructed and expressed in E.coli BL21 which provides a foundation for the further study of A30Pα-synuclein in Parkinson＇s disease.