重组人神经肽Y受体Y2融合蛋白的表达、纯化及其生物信息学分析研究

Study on expression,purification and bioinformatic analysis of recombinant human neuropeptideY Y2 receptor

目的在大肠杆菌中表达人神经肽YY2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-Y2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印迹鉴定,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Y2受体蛋白。结果经IPTG诱导含有pET28a-Y2重组质粒的DE3菌,表达出重组人Y2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Y2受体的相关生物学特性。结论重组质粒pET28a-Y2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Y2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。

Objective To express human Y2 receptor protein in E. coli,purify and identify it,and conduct bioinformatic analysis of Y2 receptor protein. Methods The recombinant plasmid pET28a-Y2 which had been well constructed and sequentially confirmed was transplanted into E. coli BL21 （ 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 Ni^2＋-NTA affinity chromatography. Then bioinformatic analysis of the Y2 receptor was conducted with the help of related online software. Results After being induced by IPTG, the DE3 with recombinant plasmid pET28a-Y2 expressed recombinant human Y2 receptor protein. Highly purified fusion protein was obtained by Ni^2＋-NTA affinity chromatography. Related biological characteristics of Y2 receptor were obtained after the online software analysis. Conclusions The recombinant plasmid pET28a-Y2 can be successfully expressed in DE3. Highly purified proteins can be obtained by Ni^2＋-NTA affinity chromatography. Y2 receptor＇s biological characteristics are predicted, which lays foundation for further studies of Y2 receptor protein＇s biological function and antibody development.