ISSN: 0973-7510

E-ISSN: 2581-690X

S. Rajagunalan , Soni Doimari and D.K. Singh
1Division of Veterinary Public Health, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh- 243 122, India.
J Pure Appl Microbiol. 2014;8(1):339-344
© The Author(s). 2014
Received: 06/08/2013 | Accepted: 20/10/2013 | Published: 28/02/2014
Abstract

Brucellosis is an important zoonotic disease transmitted to human and is also responsible for causing huge economic loss to livestock industry worldwide. Numerous candidate antigens of Brucella have been reported to induce protective effect against the organism, among which L7/L12 is an important one. In this study, the L7/L12 ribosomal gene of Brucella melitensis 16M was PCR amplified and cloned into prokaryotic expression vector pPROExHTb and the positive clones were confirmed by restriction enzyme digestion and sequencing. Recombinant clone was induced with 1mM of IPTG and the expressed protein was purified by Nickel affinity chromatography and quantified. The purified the recombinant L7/L12 protein was subjected to western blot analysis with hyperimmune serum, raised against it in rabbits and also with pooled clinical serum which revealed specific band of 16kDa confirming its antigenicity. This recombinant L7/L12 antigen can be used as protective antigen for control of brucellosis.

Keywords

Brucella melitensis, cloning, expression, L7/L12 gene

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© The Author(s) 2014. Open Access. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License which permits unrestricted use, sharing, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.