期刊文献+

结核分枝杆菌分泌蛋白MPT53基因的克隆、表达和纯化 被引量:3

Cloning, expression and purification of MPT53 of mycobacterial tuberculosis
下载PDF
导出
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。 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.
出处 《中国病原生物学杂志》 CSCD 2007年第1期14-16,共3页 Journal of Pathogen Biology
基金 上海市科委资助项目(No.05DZ22320)。
关键词 结核分枝杆菌 MPT53 克隆 表达 Mycobacteriurn tuberculosis MPT53 clone expression
  • 相关文献

参考文献6

  • 1Arati B,Kamath,Joshua Woodworth,et al.Cytolytic CD8+ T cells recognizing CFP10 are recruited to the lung after Mycobacterium tuberculosis Infection[J].Journal of Exrerimental Medicine,2004,200:1479 -1489.
  • 2Alexander C,Maue W,Ray Waters,et al.Analysis of immune responses directed toward a recombinant early secretory antigenic target sixkilodalton protein culture filtrateprotein 10 fusion protein in mycobacterium bovis-infected cattle[J].Infection and Immunity,2005,73:6659-6667.
  • 3Weldingh K,Rosenkrands I,Okkels LM,et al.assessing the serodiagnostic potential of 35 Mycobacterium tuberculosis proteins and identification of four novel serological Antigens[J].Journal of Clinical Microbiology,2005,43:57-65.
  • 4Wiker,HGS,Michell L.Cloning expression and significance of MPT53 for identification of secreted proteins of Mycobacterium tuberculosis[J].Microbial Pathogenesis,1999,26:207-219.
  • 5Johnson S,Brusasca P,Lyashchenko K,et al.Characterization of the secreted MPT53 antigen of Mycobacterium tuberculosis[J].Infection and Immunity,2001,69:5936-5939.
  • 6Hahn MH,Raman S,Anaya M,et al.The Mycobacterium tuberculosis extracytoplasmic-function sigmafactor sigl regulates polyketide synthases and secreted or membrane proteins and is required for virulence[J].Journal of Bacteriology,2005,187:7062-7071.

同被引文献15

  • 1王成勇,李益荣.3479例肺结核病人痰标本抗酸杆菌检查结果分析[J].临床肺科杂志,2004,9(6):605-606. 被引量:37
  • 2Furin JJ,Johnson JL.Recent advances in the diagnosis and management of tuberculosis.Curr Opin Pulm Med,2005,11:189-194.
  • 3Hoff ST,Abebe M,Ravn P,et al.Evaluation of mycobacterium tuberculosis--specific antibody responses in populations with different levels of exposure from Tanzania,Ethiopia,Brazil,and Denmark.Clin Infect Dis,2007,45:575-582.
  • 4Weinrieh Olsen A,van Pinxteren LA,Meng OL,et al.Protection of mice with a tuberculosis subunit vaccine based on a fusion protein of antigen 85b and esat-6.Infect Immun,2001,69:2773-2778.
  • 5Coler RN,Campos-Neto A,Ovendale P,et al.Vaccination with the T cell antigen Mtb 8.4 protects Igainst challenge with Mycobacterium tuberculosis.J Immunol,2001,166:6227-6235.
  • 6Kulshrestha A,Gupta A,Verma N,et al.Expression and purification of recombinant antigens of Mycobacterium tuberculosis for application in serodiagnosis.Protein Expr Purif,2005,44:75-85.
  • 7胡忠义.结核病免疫学诊断技术研究进展[J].中国防痨杂志,2005,27:6-7.
  • 8中国防痨协会基础专业委员会.结核病诊断实验室检验规程[M].北京:中国教育出版社,2006.13-16.
  • 9Furin J J,Johnson JL.Recent advances in the diagnosis and management of tuberculosis[J].Curt Opin Pulm Med,2005,11 (3):189-194.
  • 10Silva VMC,Kanaujia G,Gennaro ML,et al.Factors associated with humoral response to ESAT-6,38kDa and 14kDa antigens in patients with a spectrum of tuberculosis[J].Int J Tuberc Lung Dis,2003,7(5):478 -484.

引证文献3

二级引证文献16

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部