IMMUNOREACTIVE RECOMBINANT NITRATE REDUCTASE EXPRESSED BY NIR A GENE OF Mycobacterium avium sub sp paratuberculosis IS900 ELEMENT IN E.coli

C. PADMAVATHY¹*, K. VIJAYARANI², K. KUMANAN³, A. THANGAVELU⁴, A. SRITHAR^5
1Institute of Veterinary Preventive Medicine (IVPM), Ranipet, 632402, Vellore District, Tamil Nadu, India
²Department of Animal Biotechnology, Madras Veterinary College, Tamil Nadu Veterinary and Animal Sciences University, Chennai, 600 007, Tamil Nadu, India
³Centre for Animal Health Studies (CAHS), Tamil Nadu Veterinary and Animal Sciences University, Chennai, 600 051, Tamil Nadu, India
⁴Department of Veterinary Microbiology, Madras Veterinary College, Tamil Nadu Veterinary and Animal Sciences University, Chennai, 600 007, Tamil Nadu, India
⁵Department of Veterinary Microbiology, Madras Veterinary College, Tamil Nadu Veterinary and Animal Sciences University, Chennai, 600 007, Tamil Nadu, India
* Corresponding Author : kpkumarr@rediffmail.com

Received : 12-03-2018     Accepted : 16-03-2018     Published : 30-03-2018
Volume : 10     Issue : 3       Pages : 1062 - 1065
Int J Microbiol Res 10.3 (2018):1062-1065
DOI : http://dx.doi.org/10.9735/0975-5276.10.3.1062-1065

Keywords : Mycobacterium-IS900-Recombinant antigen –diagnostic candidate
Conflict of Interest : None declared
Acknowledgements/Funding : Author thankful to Prof A. Raja, Department of Animal Biotechnology, Madras Veterinary College, Chennai, 600 007, Tamil Nadu for his technical support in bioinformatics. Author thankful to Institute of Veterinary Preventive Medicine (IVPM), Ranipet, 632402, Vellore, Tamil Nadu for kindly providing bovine serum samples tested for par tuberculosis antibodies. Author also thankful to Tamil Nadu Veterinary and Animal Sciences University, Chennai, 600 051, Tamil Nadu
Author Contribution : All author equally contributed

Mycobacterium avium subsp paratuberculosis (MAP) is classified under list B transmissible organisms signifying the public health concern and needs specific early diagnosis. Due to the cross reacting antigens of environmental mycobacterium and the diverse immune responses of individuals, cocktail of antigens capable of inducing strong immune response should be identified to improve serum-based methods of detection. Insertional sequence (IS900) was specific to par tuberculosis among the mycobacterium family, and among the IS900 elements, a nir A gene at locus 4 was putative to MAP and absent from the M. avium genome. In an attempt to identify the candidate antigens, nitrate reductase gene nir A was successfully cloned and expressed in E. coli. In Western blot analysis strong immune reactivity was shown by the recombinant protein nitrate reductase with bovine anti MAP antibodies facilitates its use as a diagnostic candidate to improve the specificity of MAP diagnosis.

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