【摘要】 目的在大肠杆菌中表达人神经肽YY1受体,并对之进行纯化、鉴定及生物信息学分析。方法取已构建好且经测序确认无误的重组质粒pET28a-Y1转化大肠杆菌BL21(DE3),IPTG诱导表达融合蛋白,并经SDS-PAGE检测和Western Blot鉴定,表达产物包涵体经Ni2+-NTA亲和层析纯化。然后利用相关在线软件进行生物信息学分析Y1受体蛋白。结果经IPTG诱导含有pET28a-Y1重组质粒的DE3菌,表达出重组人Y1融合蛋白。重组蛋白经Ni2+-NTA亲和层析进行纯化后,得到了较高纯度的融合蛋白。经相关在线软件分析后获得了Y1受体的相关生物学特性。结论重组质粒pET28a-Y1在大肠杆菌DE3中成功表达,亲和层析纯化后获得较高纯度融合蛋白,并对Y1受体蛋白的生物学特征进行了预测,为进一步研究其生物学功能及其抗体的研制奠定了基础。更多还原

【Abstract】 Objective To express human Y1 receptor protein in E. coli,purify and identify it,and conduct bioinformatic analysis of Y1 receptor protein. Methods The recombinant plasmid pET28a-Y1 which had been well constructed and sequentially confirmed was transplanted into E.coliBL21 (DE3) and induced by IPTG to express fusion proteins. SDS-PAGE and Western blot were used to test and identify the expressed fusion proteins. The inclusion body of the expressed product was purified by Ni2+-NTA affinity chromatography. Then bioinformatic analysis of the Y1 receptor was conducted with the help of related online software. Results After being induced by IPTG,the DE3 with recombinant plasmid pET28a-Y1 expressed recombinant human Y1 receptor protein. Highly purified fusion protein was obtained by Ni2+-NTA affinity chromatography. Related biological characteristics of Y1 receptor were obtained after the online software analysis. Conclusions The recombinant plasmid pET28a-Y1 can be successfully expressed in DE3. Highly purified proteins can be obtained by Ni2+-NTA affinity chromatography. Y1 receptor′s biological characteristics are predicted,which lay foundation for further studies of Y1 receptor protein′s biological function and antibody development.更多还原