| 天花粉蛋白突变体TCS_(KR173-174CG)的构建、表达与纯化 |
| |
| 引用本文： | 安群星,穆士杰,张献清,陈蕤,夏爱军,张清平,陈晨,雷迎峰,黎志东,徐志凯. 天花粉蛋白突变体TCS_(KR173-174CG)的构建、表达与纯化[J]. 中国生物制品学杂志, 2006, 19(6): 557-559,563 |
| |
| 作者姓名： | 安群星 穆士杰 张献清 陈蕤 夏爱军 张清平 陈晨 雷迎峰 黎志东 徐志凯 |
| |
| 作者单位： | 第四军医大学西京医院输血科 (西安710033)第四军医大学微生物学教研室(西安710032)(安群星)，第四军医大学西京医院输血科 (西安710033)(穆士杰，张献清，陈蕤，夏爱军，张清平，陈晨)，第四军医大学微生物学教研室 (西安710032)(雷迎峰，黎志东，徐志凯) |
| |
| 摘 要： | 目的构建天花粉蛋白(TCS)突变体基因,并进行表达及纯化。方法应用计算机预测TCS分子上可能的抗原决定簇,并进行定点突变。以栝楼基因组DNA为模板,经PCR扩增突变基因,与pRSET-A表达载体连接,转化大肠杆菌BL21(DE3),经IPTG诱导表达,并对表达产物进行Ni-NTA层析纯化。结果目的蛋白在大肠杆菌中获得高效可溶性表达,表达产物经纯化后,得到均一的TCS突变体蛋白。结论已成功构建了TCS突变体基因,并获得高效表达。
|
| 关 键 词： | 天花粉蛋白 定点突变 原核表达 |
| 收稿时间： | 2006-06-19 |
| 修稿时间： | 2006-06-19 |
Construction and Expression of Trichosanthin Mutant Gene TCSKR173-174CG and Purification of Expressed Product |
| |
| AN Qun-xing ,MU Shi-jie, ZHANG Xian-qing, et al. Construction and Expression of Trichosanthin Mutant Gene TCSKR173-174CG and Purification of Expressed Product[J]. Chinese Journal of Bilogicals, 2006, 19(6): 557-559,563 |
| |
| Authors: | AN Qun-xing MU Shi-jie ZHANG Xian-qing et al |
| |
| Abstract: | Objective To construct and express a trichosanthin(TCS) gene mutant and purify the expressed product.Methods Predict the potential antigenic determinant on TCS molecule by computer modeling and induce site-directed mutation.Amplify gene mutant TCS KR173-174CG by PCR using the genomic DNA of Trichosanthes kirilowii as a template and insert into expression vector pRSET-A,then transform to E.coli BL21(DE3) for expression under induction of IPTG.Purify the expressed product by Ni-NTA affinity column chromatography.Results The target protein in a soluble form was successfully expressed in E.coli.Homogenous TCS mutant protein was obtained after purification of expressed product.Conclusion TCS mutant gene TCS KR173-174CG was successfully constructed and expressed. |
| |
| Keywords: | Trichosanthin Site-directed mutation Prokaryotic expression |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
|
