Antigenicity of recombinant L7/L12 in patients with brucellosis

authors:

avatar Hamid Abtahi ORCID , * , avatar Ali hatef Salmanian

Koomesh: Vol.13, issue 2; 166-171
published online: May 25, 2012
article type: Research Article
received: January 01, 2011
accepted: July 30, 2011

how to cite: Abtahi H, Salmanian A H. Antigenicity of recombinant L7/L12 in patients with brucellosis. koomesh. 2012;13(2):e152504. 

Abstract

  Introduction: Immune response to recombinant L7/L12, in addition to protective role, may show its importance in detection Brucellosis tests. The aim of this study was to examine antigenicity of recombinant L7/L12 from Brucella abortus by Brucellosis human sera.   Material and Methods : We amplified L7/L12 gene by polymerase chain reaction (PCR) method and sub- cloned to prokaryotic expression vector pET28a. Escherichia coli BL21-DE3-plySs was transformed with pET28a-L7/L12 and gene expression was induced by IPTG. Recombinant L7/L12 was further analyzed by Western Blot. Sera reactivity of five infected individual were further analyzed against the recombinant L7/L12 protein.   Results: The sequencing result was confirmed by Sanger method and it was the same as L7/L12 gene. Escherichia coli BL21 (DE3) pLysS was transformed with pET28a-L7/L12 and gene expression was induced by IPTG. The expressed protein was purified by affinity chromatography by Ni-NTA resin. The data also indicated that L7/L12 protein from Brucella abortus recognized by patient sera.   Conclusion: Our data showed that recombinant L7/L12 protein can be produced by pET28a in Escherichia coli. This protein was recognized by sera in infected human as an antigen. Therefore, recombinant L7/L12 has same epitopes with natural form of this antigen. Recombinant L7/L12 also seems to be a promising antigen for protection and serologic diagnosis of Human brucellosis.