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Expression and purification of Mycobacterium tuberculosis ESAT-6 and MPT64 fusion protein and its immunoprophylactic potential in mouse model.
- Source :
-
Protein expression and purification [Protein Expr Purif] 2008 Jun; Vol. 59 (2), pp. 189-96. Date of Electronic Publication: 2007 Dec 15. - Publication Year :
- 2008
-
Abstract
- 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.
- Subjects :
- Animals
Antigens, Bacterial isolation & purification
Antigens, Bacterial therapeutic use
Bacterial Proteins isolation & purification
Bacterial Proteins therapeutic use
Blotting, Western
Chromatography, Affinity
Cloning, Molecular
Disease Models, Animal
Escherichia coli genetics
Genetic Vectors
Humans
Interferon-gamma metabolism
Interleukin-12 metabolism
Lymphocyte Activation
Mice
Mice, Inbred C57BL
Mycobacterium tuberculosis isolation & purification
Nitrilotriacetic Acid chemistry
Protein Folding
Recombinant Proteins isolation & purification
Recombinant Proteins therapeutic use
Tuberculosis immunology
Tuberculosis Vaccines isolation & purification
Tuberculosis Vaccines therapeutic use
Antigens, Bacterial biosynthesis
Bacterial Proteins biosynthesis
Mycobacterium tuberculosis immunology
Recombinant Proteins biosynthesis
Tuberculosis prevention & control
Tuberculosis Vaccines biosynthesis
Subjects
Details
- Language :
- English
- ISSN :
- 1096-0279
- Volume :
- 59
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Protein expression and purification
- Publication Type :
- Academic Journal
- Accession number :
- 18378164
- Full Text :
- https://doi.org/10.1016/j.pep.2007.11.016