3BP2多克隆抗体的制备及其与SHP-1之间的相互关系

发布时间：2018-01-10 09:00

本文关键词：3BP2多克隆抗体的制备及其与SHP-1之间的相互关系　出处：《吉林大学》2006年硕士论文　论文类型：学位论文

【摘要】：为了研究人的适配子蛋白3BP2(又称作SH3BP2)在细胞信号转导中的生物学功能,本研究以3BP2的全长cDNA为模板,克隆构建了原核融合蛋白表达载体pGex2T-3BP2质粒,转化大肠杆菌DE3-plysS,进行融合蛋白质GST-3BP2的高效表达,并利用Gluthione交联的Sephorose-4B亲和层析柱纯化该融合蛋白质,制备了GST-3BP2。GST-3BP2免疫兔得到这个融合蛋白抗血清,通过抗体的负选择和正选择亲和层析柱进行纯化,制备了抗3BP2兔的多克隆抗体。利用免疫沉淀(Immunoprecipitation)和Western blot等生物学研究手段,在鼠脾中检测到了3BP2与SHP1的相互作用。3BP2作为酪氨酸磷酸激酶(PTK)Lyn,Syk的底物参与到与酪氨酸磷酸酶(PTP)SHP-1的信号通路中,这对于阐明与3BP2相关的TCR,BCR和FcεRI介导的信号通路,深入研究PTP和PTK之间的关系,为3BP2相关疾病过敏性疾病和巨颌症药物靶点的研究奠定了理论基础。
[Abstract]:In order to study the biological function of human aptamer protein 3BP2( also known as SH3BP2) in cell signal transduction, the full-length cDNA of 3BP2 was used as a template in this study. The prokaryotic fusion protein expression vector pGex2T-3BP2 plasmid was constructed and transformed into E. coli DE3-plysSto express the fusion protein GST-3BP2. The fusion protein was purified by Gluthione crosslinked Sephorose-4B affinity chromatography. The antiserum of the fusion protein was prepared from rabbits immunized with GST-3BP2.GST-3BP2 and purified by negative and positive selection affinity chromatography. The polyclonal antibody against 3BP2 rabbit was prepared, and immunoprecipitation and Western blot were used to study it. The interaction between 3BP2 and SHP1 was detected in mouse spleen. 3BP2 was used as tyrosine phosphokinase (PTK) Lyn. The substrates of Syk are involved in the signal pathway with tyrosine phosphatase PTPP-SHP-1, which is helpful to elucidate the signal pathway mediated by 3BP2, TCR-BCR and FC 蔚 RI. The study of the relationship between PTP and PTK lays a theoretical foundation for the study of drug targets for 3BP2 related diseases, allergic diseases and megalomaxillary diseases.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R392

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