Brucellosis is a well-known infectious disease among domestic animals caused by Brucella bacterium. OMP25 is an outer membrane protein of Brucella which plays an important role in immunogenicity of cells. Moreover, Brucella Lumazine Synthase (BLS) as another main antigen can be used as an adjuvant when covalently attached to a foreign antigen. In the current study, designing and production of a chimeric construct as a primary step to stimulate the immune system against Brucella infection is investigated. The construct was amplified by specific primers using SOE-PCR technique. The amplified construct was cloned in pTZ57R/T vector and transformed into E. coli TOP10F’ as cloning host. Then, the construct was subcloned in pET-32a (+) vector followed by transforming into E. coli BL21 (DE3) as expression host. Our results demonstrated OMP25-BLS with 1163 bp was successfully cloned and expressed in the expression host. Results were confirmed using sequencing, SDS-PAGE and western blotting which correctly showed 59 KD protein band of OMP25-BLS. According to our results this construct could be proposed to investigate as a new subunit vaccine candidate in order to stimulate immune system against Brucellosis in future studies.

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