摘要
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 neuropeptideY （NPY） protein in E. coli, purify and identify it, and conduct bioinformatic analysis of NPY protein. Methods The recombinant plasmid pET28a-NPY which had been well constructed and sequentially confirmed was transplanted into E. coli BL21 （DE3） and induced by IPTC to express fusion proteins. SDS-PAGE and Western blot were used to test and i- dentify the expressed fusion proteins. The inclusion body of the expressed product was purified by Ni2 ＋ -NTA affinity chromatography. Then bioinformatie analysis of the NPY protein was conducted with the help of related online software. Results After being induced by IPTG,the DE3 with recombinant plasmid pET28a-NPY expressed recombinant human NPY protein. Highly purified NPY fusion protein was obtained by Ni2＋ -NTA affinity chromatography. Related biological characteristics of NPY were obtained after the online software analysis. Conclusions The recombinant plasmid pET28a-NPY can be successfully expressed in DE3. Highly purified proteins can be obtained by Ni2＋ -NTA affinity chromatography. NPY＇s biological characteristics are predicted, which lays foundation for further studies of NPY＇s biological function and antibody development.
出处
《中国老年学杂志》
CAS
CSCD
北大核心
2009年第17期2198-2202,共5页
Chinese Journal of Gerontology
关键词
人神经肽Y
融合蛋白
包涵体
纯化
生物信息学
Neuropeptide Y
Fusion protein
Inclusion body
Purification
Bioinformatics
