双磷酸化His-PZR蛋白的表达与分离纯化

Expression and purification of bi-phosphorylated His-PZR protein

目的获得纯化的双磷酸化His-PZR蛋白。方法本试验采用His-PZR与C-Src重组原核表达载体共同转化E.coliBL21菌株的方法,通过IPTG诱导表达,Ni-NTA纯化及HPLC分离纯化等技术,获得双磷酸化的His-PZR蛋白。结果成功表达了His-PZR蛋白,约占全菌体蛋白的8.5%,相对分子质量约为10 000;并分离纯化出双磷酸化的His-PZR蛋白。结论双磷酸化His-PZR蛋白的表达和纯化,为进一步研究His-PZR与SHP蛋白间的作用机制及其在信号转导过程中的功能奠定了基础。

Objective To express and purify the bi-phosphorylated His-PZR proteins.Methods The His-PZR and C-Src recombined prokaryotic expression plasmids were co-transformed to the E.Coli BL21.IPTG induction,Ni-NTA purification,and HPLC separation were utilized to get the bi-phosphorylated His-PZR proteins.Results The His-PZR protein had successfully been expressed,which was 8.5% of the total bacterial protein in E.Coli BL21,and possessed the relative molecular mass of 10 000.After Ni-NTA purification and HPLC separa...