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 |
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引用本文: | 丁克祥,董萍,郑永晨,杨永鹏,朱晓亮,韩晋云,单志新,丁宇,丁振华.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[J].中国老年学杂志,2009,29(19). |
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作者姓名: | 丁克祥 董萍 郑永晨 杨永鹏 朱晓亮 韩晋云 单志新 丁宇 丁振华 |
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作者单位: | 1. 南方医科大学科研部,广东,广州,510515 2. 大连董萍美容整形医院 3. 吉林大学附属第二临床医院 4. 南方医科大学附属南方医院 5. 南方医科大学研究生学院 6. 南方医科大学附属华南医院 7. 华中科技大学同济医学院附属同济医院 8. 南方医科大学公共卫生与热带医学学院 |
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摘 要: | 目的 在大肠杆菌中表达人神经肽Y Y2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Ni~(2+)-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Ni~(2+)-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.
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关 键 词: | 人神经肽Y Y2受体 融合蛋白 包涵体 纯化 生物信息学 |
Study on expression, purification and bioinformatic analysis of recombinant human neuropeptideY Y2 receptor |
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Abstract: | 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 Ni2+-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 Ni2+-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. |
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Keywords: | NeuropeptideY Y2 receptor Fusion protein Inclusion body Purification Bioinformatics |
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