Expression and purification of Mycobacterium tuberculosis ESAT-6 and MPT64 fusion protein and its immunoprophylactic potential in mouse model.
The completion of Mycobacterium tuberculosis genome sequence has opened a new way for the identification and characterization of bacterial antigens, such as ESAT-6, CFP10, MPT64, and Ag85 complex, which are helpful for tuberculosis control. In this work, genes of ESAT-6 and MPT64 were fused and expressed in Escherichia coli in form of inclusion bodies with a histidine tag. The expressed fusion protein was purified by nitrilotriacetic acid (Ni-NTA) affinity chromatography under denaturing conditions, and the yield was 18mg/L of culture. In mice, the purified ESAT-6-MPT64 fusion protein elicited stronger humoral response, greater splenic lymphocyte stimulated index, and higher levels of IFN-gamma and IL-12 production than that of the single MPT64 inoculation group, and rendered modest protection on the experimental tuberculosis mouse models. In short, the ESAT-6-MPT64 fusion protein might be a potential candidate vaccine for tuberculosis.
Bai
Yinlan
Y
Department of Microbiology, Fourth Military Medical University, 17th Changle West Road, Xi'an 710032, Shannxi Province, China.
Xue
Ying
Y
Gao
Hui
H
Wang
Limei
L
Ding
Tianbing
T
Bai
Wentao
W
Fan
Ailin
A
Zhang
Jianfang
J
An
Qunxing
Q
Xu
Zhikai
Z
D000818:Animals; D000942:Antigens, Bacterial / Q000096:biosynthesis* / Q000302:isolation & purification / Q000627:therapeutic use; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000302:isolation & purification / Q000627:therapeutic use; D015153:Blotting, Western; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004195:Disease Models, Animal; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D018664:Interleukin-12 / Q000378:metabolism; D008213:Lymphocyte Activation; D051379:Mice; D008810:Mice, Inbred C57BL; D009169:Mycobacterium tuberculosis / Q000276:immunology* / Q000302:isolation & purification; D009571:Nitrilotriacetic Acid / Q000737:chemistry; D017510:Protein Folding; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000302:isolation & purification / Q000627:therapeutic use; D014376:Tuberculosis / Q000276:immunology / Q000517:prevention & control*; D032581:Tuberculosis Vaccines / Q000096:biosynthesis* / Q000302:isolation & purification / Q000627:therapeutic use
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; C093276:ESAT-6 protein, Mycobacterium tuberculosis; C080865:MPB64 protein, Mycobacterium; D011994:Recombinant Proteins; D032581:Tuberculosis Vaccines; D018664:Interleukin-12; D007371:Interferon-gamma; D009571:Nitrilotriacetic Acid