| GST-p16融合蛋白的表达、纯化及鉴定 |
| |
| 引用本文： | 史从宁,吕凤祥,王天英,郭卓维,傅国辉. GST-p16融合蛋白的表达、纯化及鉴定[J]. 哈尔滨医科大学学报, 2004, 38(3): 215-217 |
| |
| 作者姓名： | 史从宁 吕凤祥 王天英 郭卓维 傅国辉 |
| |
| 作者单位： | 哈尔滨医科大学,病理生理学教研室,黑龙江,哈尔滨,150086;哈尔滨医科大学,病理生理学教研室,黑龙江,哈尔滨,150086;哈尔滨医科大学,病理生理学教研室,黑龙江,哈尔滨,150086;哈尔滨医科大学,病理生理学教研室,黑龙江,哈尔滨,150086;哈尔滨医科大学,病理生理学教研室,黑龙江,哈尔滨,150086 |
| |
| 基金项目： | 国家自然科学基金重点项目资助 (3 0 2 3 0 160 ) |
| |
| 摘 要： | 目的利用GST融合基因表达系统表达、纯化及鉴定GST-p16融合蛋白。方法以质粒pM-p16为模板，扩增出p16基因片段，将其克隆至大肠杆菌表达载体pGEX-6P-1，将所构建的重组质粒pGEX-p16转化大肠杆菌B121并诱导表达，采用GST蛋白纯化系统进行纯化，所得产物进行SDS-PAGE及Westernblot鉴定。结果大肠杆菌细胞经诱导高效表达出约42kD蛋白，其分子量与GST-p16融合蛋白相符。Westernblot结果显示该蛋白能够被p16．抗体特异性识别。结论本实验在大肠杆菌表达系统中高效表达了有活性的GSTipl6融合蛋白并可用于以后的实验研究。
|
| 关 键 词： | p16 GST融合基因表达系统 Westem blot |
| 文章编号： | 1000-1905(2004)03-0215-03 |
| 修稿时间： | 2004-02-13 |
Expression, purification and identification of recombinant GST-p16 fusion protein |
| |
| SHI Cong ning,LU Feng xiang,WANG Tian ying,et al. Expression, purification and identification of recombinant GST-p16 fusion protein[J]. Journal of Harbin Medical University, 2004, 38(3): 215-217 |
| |
| Authors: | SHI Cong ning LU Feng xiang WANG Tian ying et al |
| |
| Abstract: | Objective To express,purify and identify recombinant GST p16 fusion protein using GST gene fusion system.Methods The cDNA of p16 was amplified from plasmid pM p16 and cloned into pGEX 6P 1 expression vector.The recombinant plasmid pGEX p16 was expressed in E.coli BL21 cells and the products were purified by GST purifying system.The specific expression was identified by Western blot.Results The resolved GST p16 fusion protein on 15% SDS PAGE showed a major band at position of 42kD and the fusion protein was recognized by anti p16 antibody on PVDF membrane.Conclusion The recombinant GST p16 fusion protein is expressed in E.coli cells efficiently,and the purified GST p16 fusion protein can be used in further study. |
| |
| Keywords: | p16 protein GST gene fusion system Western blot |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
|
