Cloning, expression and purification of Mycobacterium tuberculosis ESAT-6 and CFP-10 antigens

被引：0
|
作者：
Mahmoudi, Shima [1 ]
Mamishi, Setareh [1 ,2 ]
Ghazi, Mona [3 ]
Sadeghi, Reihaneh Hosseinpour [1 ]
Pourakbari, Babak [1 ]
机构：
[1] Univ Tehran Med Sci, Pediat Infect Dis Res Ctr, Tehran, Iran
[2] Univ Tehran Med Sci, Children Med Ctr Hosp, Sch Med, Dept Infect Dis, Tehran, Iran
[3] Shahid Beheshti Univ Med Sci, Sch Med, Dept Microbiol, Tehran, Iran
来源：
IRANIAN JOURNAL OF MICROBIOLOGY | 2013年 / 5卷 / 04期
关键词：
M; tuberculosis; ESAT-6; CFP-10;
D O I：
暂无
中图分类号：
Q93 [微生物学];
学科分类号：
071005 ; 100705 ;
摘要：
Background and Objectives: ESAT-6 (6-kDaearly secretory antigenic target) and CFP-10 (10-kDa culture filtrate protein) have been described as dominant antigens recognized by T-cells and considered as virulence factors in Mycobacterium tuberculosis. The aim of this study was to clone, express and purify recombinant ESAT-6 and CFP-10 proteins of M. tuberculosis in soluble form. Materials and Methods: ESAT-6 and CFP-10 genes were amplified by PCR, cloned into pET32a (+) vector, and overexpressed using isopropyl-beta-D-thiogalactopyranoside in E. coli BL21 (DE3). ESAT-6 andCFP-10 proteins were purified by Ni-NTA affinity chromatography and were detected by anti-ESAT-6 and anti-CFP10 antibodies. Results: ESAT-6 andCFP-10 genes were successfully expressed and purified. Anti-ESAT-6 and anti-CFP-10 antibodies were produced after induction of immunization against purified ESAT-6 andCFP-10 proteins in rabbit. Conclusion: In this study, we cloned, expressed and purified sufficient amounts of ESAT-6 andCFP-10 and it would be tested for the development of diagnostic kit for M. tuberculosis in future.
引用
收藏
页码：374 / 378
页数：5
相关论文
共 50 条
  • [21] Rapid Identification of the Mycobacterium tuberculosis Complex by Combining the ESAT-6/CFP-10 Immunochromatographic Assay and Smear Morphology
    Shen, Gwan-Han
    Chiou, Chien-Shun
    Hu, Shiau-Ting
    Wu, Kun-Ming
    Chen, Jiann-Hwa
    JOURNAL OF CLINICAL MICROBIOLOGY, 2011, 49 (03) : 902 - 907
  • [22] Novel monoclonal antibodies to ESAT-6 and CFP-10 antigens for ELISA-based diagnosis of pleural tuberculosis
    Feng, T-T.
    Shou, C-M.
    Shen, L.
    Qian, Y.
    Wu, Z-G.
    Fan, J.
    Zhang, Y-Z.
    Tang, Y-W.
    Wu, N-P.
    Lu, H-Z.
    Yao, H-P.
    INTERNATIONAL JOURNAL OF TUBERCULOSIS AND LUNG DISEASE, 2011, 15 (06) : 804 - 810
  • [23] Factors associated with differential T cell responses to antigens ESAT-6 and CFP-10 in pulmonary tuberculosis patients
    Liu, Shengsheng
    Wu, Meiying
    Ertai, A.
    Wu, Shucai
    Geng, Shujun
    Li, Zhihui
    Li, Mingwu
    Li, Liang
    Pang, Yu
    Kang, Wanli
    Tang, Shenjie
    MEDICINE, 2021, 100 (08) : E24615
  • [24] ESAT-6 and CFP-10: What is the diagnosis? Authors' reply
    Geluk, A
    Ottenhoff, THM
    Arend, SM
    Andersen, P
    Doherty, TM
    INFECTION AND IMMUNITY, 2002, 70 (11) : 6510 - 6511
  • [25] Prospective evaluation of a whole-blood test using Mycobacterium tuberculosis-specific antigens ESAT-6 and CFP-10 for diagnosis of active tuberculosis
    Ravn, P
    Munk, ME
    Andersen, ÅB
    Lundgren, B
    Lundgren, JD
    Nielsen, LN
    Kok-Jensen, A
    Andersen, P
    Weldingh, K
    CLINICAL AND DIAGNOSTIC LABORATORY IMMUNOLOGY, 2005, 12 (04) : 491 - 496
  • [26] Structure and function of the complex formed by the tuberculosis virulence factors CFP-10 and ESAT-6
    Renshaw, PS
    Lightbody, KL
    Veverka, V
    Muskett, FW
    Kelly, G
    Frenkiel, TA
    Gordon, SV
    Hewinson, RG
    Burke, B
    Norman, J
    Williamson, RA
    Carr, MD
    EMBO JOURNAL, 2005, 24 (14): : 2491 - 2498
  • [27] The CFP-10/ESAT-6 complex of Mycobacterium tuberculosis potentiates the activation of murine macrophages involvement of IFN-γ signaling
    Si Guo
    Lang Bao
    Zi Fang Qin
    Xin Xin Shi
    Medical Microbiology and Immunology, 2010, 199 : 129 - 137
  • [28] Detection of BAL Cell Responses to CFP-10 and ESAT-6 in Healthy Mycobacterium tuberculosis-Infected Individuals.
    Walrath, J. R.
    Lee, H. I.
    Silver, R. F.
    AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE, 2009, 179
  • [29] Comparison of different immunization routes on the immune responses induced by Mycobacterium tuberculosis ESAT-6/CFP-10 recombinant protein
    Namvarpour, Mozhdeh
    Tebianian, Majid
    Mansouri, Reza
    Ebrahimi, Seyyed Mahmoud
    Kashkooli, Shiva
    BIOLOGICALS, 2019, 59 : 6 - 11
  • [30] Individual RD1-region genes are required for export of ESAT-6/CFP-10 and for virulence of Mycobacterium tuberculosis
    Guinn, KM
    Hickey, MJ
    Mathur, SK
    Zakel, KL
    Grotzke, JE
    Lewinsohn, DM
    Smith, S
    Sherman, DR
    MOLECULAR MICROBIOLOGY, 2004, 51 (02) : 359 - 370
12345