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目的在大肠杆菌中表达人神经肽Y,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-NPY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NPY蛋白。结果经IPTG诱导含有pET28a-NPY重组质粒的DE3菌,表达出重组人NPY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NPY的相关生物学特性。结论重组质粒pET28a-NPY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NPY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 neuropeptide Y in Escherichia coli and to purify, identify and bioinformatically analyze it. Methods The constructed recombinant plasmid pET28a-NPY was transformed into E.coli BL21 (DE3) and induced by IPTG. The fusion protein was induced by IPTG and identified by SDS-PAGE and Western blotting. The inclusion bodies were expressed by Ni2 + -NTA And chromatographic purification. Then use the relevant online software for bioinformatics analysis of NPY protein. Results Recombinant human NPY fusion protein was expressed by IPTG induction of DE3 containing recombinant plasmid pET28a-NPY. The recombinant protein was purified by Ni2 + -NTA affinity chromatography, and the fusion protein of higher purity was obtained. The related biological characteristics of NPY were obtained after the related online software analysis. Conclusion The recombinant plasmid pET28a-NPY was successfully expressed in E. coli DE3. The fusion protein was purified by affinity chromatography and the biological characteristics of NPY protein were predicted. In order to further study its biological function and antibody Research and laid a foundation.