|
|||||
|
|
| 结核分枝杆菌分泌蛋白MPT53基因的克隆、表达和纯化 | |
| 引用本文： | 唐宇龙,杨华,刘湘新,秦莲花,柳彬,胡忠义.结核分枝杆菌分泌蛋白MPT53基因的克隆、表达和纯化[J].中国病原生物学杂志,2007,2(1):14-16. |
| 作者姓名： | 唐宇龙 杨华 刘湘新 秦莲花 柳彬 胡忠义 |
| 作者单位： | 1. 上海市结核重点实验室,上海,200433;湖南农业大学动物医学院,湖南长沙,410128 2. 上海市结核重点实验室,上海,200433 3. 湖南农业大学动物医学院,湖南长沙,410128 |
| 摘 要： | 目的构建结核杆菌分泌蛋白MPT53原核表达载体并进行表达和纯化。方法以结核分枝杆菌标准株H37Rv基因组DNA为模板扩增MPT53基因，产物经纯化回收后与载体pMD-T连接转化，酶切鉴定，克隆到pET32a原核表达载体，测序鉴定插入序列完全正确者转化大肠埃希菌BL21，诱导表达MPT53融合蛋白，亲和层析纯化后，进行SDS-PAGE电泳鉴定。结果成功构建PET32a-MPT53重组质粒，转化BL21后经IPGT诱导成功表达分子质量单位为36ku的MPT53蛋白，与理论值相符。结论成功表达结核分枝杆菌MPT53蛋白，为其应用打下了基础。 |
| 关 键 词： | 结核分枝杆菌 克隆 表达 |
| 文章编号： | 1673-5234（2007）01-0014-03 |
| 收稿时间： | 2006-09-26 |
| 修稿时间： | 2006-09-262006-12-21 |
Cloning, expression and purification of MPT53 of mycobacterial tuberculosis |
|
| TANG Yu-long,YANG Hua,LIU Xiang-xin,QIN Lian-hua,LIU Bin,HU Zhong-yi.Cloning, expression and purification of MPT53 of mycobacterial tuberculosis[J].Journal of Pathogen Biology,2007,2(1):14-16. | |
| Authors: | TANG Yu-long YANG Hua LIU Xiang-xin QIN Lian-hua LIU Bin HU Zhong-yi |
| Institution: | 1. Shanghai TB Key Laboratory, Shanghai 200433, China ; 2. College of Veterinary Medicine, Hunan Agricultural University |
| Abstract: | Objective To construct a expression vector of MPT53 of mycobacterium tuberculosis and identify and purify the protein in the E. coli. Methods The gene encoding protein MPT53 was amplified from mycobacterium tuberculosis H37RV chromosomal DNA by using PCR, then cloned into pMD-T vector after identifying, then cloned pET32a expressing vector, transformed into E. coli BL21. 13caterial lyastes prepared from IPTG induced cultures were loading SDS-PAGE. Results Recombinant plasmid PET32a-MPT53 was constructed, and 36 ku MPT53 protein was expressed, which consistent with the theoretical value. Conclusion MPT53 protein expressed successfully and lays the foundation for its application. |
| Keywords: | MPT53 |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |