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Showing 2 results for Mohabati Mobarez
Sayeneh Khodadadi, Ashraf Mohabati Mobarez, Naser Harzandi , Bahman Tabaraei , Nima Khoramabadi, Amir Bakhtiyari , Hanyieh Aghababa ,
Volume 12, Issue 1 (spring 2012)
Abstract
Background & Objectives: The identification of Brucella spp. antigens with the capacity to elicit a protective immune response is of the great interest for the researchers. So, characterization and assessment of diverse antigens of Brucella need to be evaluated. In this study, we report the cloning and expression of the gene coding for 31 KDa OMP (OMP31) of Brucella melitensis 16M. Methods: Brucella melitensis Omp31 gene was amplified with specific primers, cloned into pJET1/2 and subsequently subcloned in pET28a (+) vector. Both these recombinant plasmids were sequenced and then after, expression of recombinant protein was induced by 1mM IPTG. Western blot analysis was also performed by polyclonal rabbit antiserum. Results: Omp31 successfully was cloned in both plasmid vectors. The recombinant Omp31 was expressed in E.coli host and purified with significant yield. Western blot results along with those of sequencing ensured accurate production of recombinant omp31 and retaining of its partial epitopes. Conclusion: Our results show that, an expression host such as E. coli is suitable for omp31 production.
Farshad Nojoomi, Ashraf Mohabati Mobarez, Ali Hatef Salmanian, Seiyed Davar Siadat, Nima Khoramabadi, Haniyeh Aghababa,
Volume 12, Issue 4 (winter 2012)
Abstract
Background & Objectives: Haemophilus influenzae is a Gram-negative bacterium that is part of the normal nasopharyngeal flora of most humans. H. influenzae strains were defined in two categories: encapsulated (typeable) and non-capsulated (nontypeable) strains. Outer membrane protein P6 is a highly conserved and stable protein in the outer membrane of both encapsulated and non-capsulated strains of H. influenzae. As an immunogen, P6 protein is a potential candidate vaccine against H. influenzae strains. The aim of this study was to produce recombinant protein P6 as a carrier protein for production of conjugate vaccines. Methods: The sequence (324 bp) coding P647-153 protein of H. influenzae was successfully cloned in pJET1.2 and subsequently in pET28a (+). Expression of the recombinant protein was induced with 1mM IPTG. Recombinant P647-153 was purified through dissolving inclusions in 8M urea buffer, absorbing to Ni-NTA resins, washing by buffers with decreasing urea concentration and finally eluted in Imidazole solution. Imidazole was removed by dialysis against PBS (pH 7.4). The recombinant p6 47-153 was confirmed by western blot analysis using rabbit anti H. influenzae polyclonal antiserum. Results: The recombinant P647-153 was successfully expressed in E. coli BL21 (DE3) and purified (4 mg /lit of broth culture). The immunoblotting showed that recombinant P647-153 conserved its native antigenic structure. Conclusion: Western blot results, along with that of sequencing, confirmed accurate production of recombinant P647-153 and partially retention of its conformational epitopes.
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