论文部分内容阅读
目的应用免疫共沉淀技术,验证新基因AngRem104和糖皮质激素受体特异延伸因子(GR-EF)蛋白在哺乳动物细胞中的相互作用,为进一步研究AngRem104的生理功能奠定基础。方法构建AngRem104-pcDNA3.1/V5-His和GR-EF-pCMV/c-myc融合表达载体,共转染HEK293T细胞。分别用小鼠抗人c-myc单克隆抗体和小鼠抗人V5-HRP抗体进行免疫沉淀和免疫印迹,以验证AngRem104和GR-EF蛋白间的相互作用。结果AngRem104-V5融合蛋白能够在c-myc抗体的免疫沉淀物中检测出来;在V5抗体的免疫沉淀物中也能检测到GR-EF-c-myc融合蛋白。结论新基因AngRem104和GR-EF蛋白在哺乳动物细胞中存在直接相互作用。
Objective To investigate the interaction between AngRem104 and glucocorticoid receptor-specific elongation factor (GR-EF) protein in mammalian cells by co-immunoprecipitation technique, and to lay the foundation for further study on the physiological function of AngRem104. Methods The fusion protein of AngRem104-pcDNA3.1 / V5-His and GR-EF-pCMV / c-myc was constructed and transfected into HEK293T cells. Immunoprecipitation and immunoblotting were performed with mouse anti-human c-myc monoclonal antibody and mouse anti-human V5-HRP antibody respectively to verify the interaction between AngRem104 and GR-EF protein. Results AngRem104-V5 fusion protein could be detected in the immunoprecipitates of c-myc antibody. GR-EF-c-myc fusion protein could also be detected in immunoprecipitates of V5 antibody. Conclusion The new gene AngRem104 and GR-EF protein have direct interaction in mammalian cells.