Design and Production of a Novel Recombinant Chimeric IL2-Omp31 Antigen against Brucella Infection
Brucellosis is a zoonotic disease in human and animals. Brucella melitensis is one of the most pathogenic species of Brucella in goat and sheep. Omp31 is an outer membrane protein of Brucella that acts as an immunogenic protein. Cytokines are glycoproteins with low molecular weight that play the rol...
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| Format: | Article |
| Language: | English |
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Razi Vaccine and Serum Research Institute
2018-09-01
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| Series: | Archives of Razi Institute |
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| Online Access: | http://archrazi.areeo.ac.ir/article_116404_4286f2e4b17f23e95d54e0bb1644ae65.pdf |
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| author | M. Naghavi M. H. Sekhavati M. Tahmoorespur M. R. Nassiri |
| author_facet | M. Naghavi M. H. Sekhavati M. Tahmoorespur M. R. Nassiri |
| author_sort | M. Naghavi |
| collection | DOAJ |
| description | Brucellosis is a zoonotic disease in human and animals. Brucella melitensis is one of the most pathogenic species of Brucella in goat and sheep. Omp31 is an outer membrane protein of Brucella that acts as an immunogenic protein. Cytokines are glycoproteins with low molecular weight that play the role of an immune adjuvant and regulate immune responses. Interleukin-2 is one of the most important cytokines, which are secreted by the white blood cells and involved in T cell immune responses. In the present study, a chimeric Omp31-Interleukin2 recombinant protein was generated by means of genetic engineering techniques. This chimeric coding sequence was amplified by using specific primers and using Splicing Overlap Extension (SOE) PCR technique. The fusion of the two mentioned proteins was accomplished using a rigid linker. The generated chimeric IL2-Omp31 fragment was TA cloned, and then subcloned into pEt22b vector as an expression vector. The chimeric protein was successfully expressed in E. coli BL21 (DE3) and confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and also Western-blotting analysis. Finally, in order to assess the antigenic features of the recombinant chimeric IL2-Opm31 protein, its secondary structure and antigenicity were predicted in silico. |
| first_indexed | 2024-12-11T14:54:39Z |
| format | Article |
| id | doaj.art-a78ae19380b84f30a70079876dcd24ee |
| institution | Directory Open Access Journal |
| issn | 0365-3439 2008-9872 |
| language | English |
| last_indexed | 2024-12-11T14:54:39Z |
| publishDate | 2018-09-01 |
| publisher | Razi Vaccine and Serum Research Institute |
| record_format | Article |
| series | Archives of Razi Institute |
| spelling | doaj.art-a78ae19380b84f30a70079876dcd24ee2022-12-22T01:01:20ZengRazi Vaccine and Serum Research InstituteArchives of Razi Institute0365-34392008-98722018-09-0173319920610.22092/ari.2017.110504.1131116404Design and Production of a Novel Recombinant Chimeric IL2-Omp31 Antigen against Brucella InfectionM. Naghavi0M. H. Sekhavati1M. Tahmoorespur2M. R. Nassiri3Department of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, IranDepartment of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, IranDepartment of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, IranDepartment of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, IranBrucellosis is a zoonotic disease in human and animals. Brucella melitensis is one of the most pathogenic species of Brucella in goat and sheep. Omp31 is an outer membrane protein of Brucella that acts as an immunogenic protein. Cytokines are glycoproteins with low molecular weight that play the role of an immune adjuvant and regulate immune responses. Interleukin-2 is one of the most important cytokines, which are secreted by the white blood cells and involved in T cell immune responses. In the present study, a chimeric Omp31-Interleukin2 recombinant protein was generated by means of genetic engineering techniques. This chimeric coding sequence was amplified by using specific primers and using Splicing Overlap Extension (SOE) PCR technique. The fusion of the two mentioned proteins was accomplished using a rigid linker. The generated chimeric IL2-Omp31 fragment was TA cloned, and then subcloned into pEt22b vector as an expression vector. The chimeric protein was successfully expressed in E. coli BL21 (DE3) and confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and also Western-blotting analysis. Finally, in order to assess the antigenic features of the recombinant chimeric IL2-Opm31 protein, its secondary structure and antigenicity were predicted in silico.http://archrazi.areeo.ac.ir/article_116404_4286f2e4b17f23e95d54e0bb1644ae65.pdfBrucella melitensisInterleukinOmp31Cytokines |
| spellingShingle | M. Naghavi M. H. Sekhavati M. Tahmoorespur M. R. Nassiri Design and Production of a Novel Recombinant Chimeric IL2-Omp31 Antigen against Brucella Infection Archives of Razi Institute Brucella melitensis Interleukin Omp31 Cytokines |
| title | Design and Production of a Novel Recombinant Chimeric IL2-Omp31 Antigen against Brucella Infection |
| title_full | Design and Production of a Novel Recombinant Chimeric IL2-Omp31 Antigen against Brucella Infection |
| title_fullStr | Design and Production of a Novel Recombinant Chimeric IL2-Omp31 Antigen against Brucella Infection |
| title_full_unstemmed | Design and Production of a Novel Recombinant Chimeric IL2-Omp31 Antigen against Brucella Infection |
| title_short | Design and Production of a Novel Recombinant Chimeric IL2-Omp31 Antigen against Brucella Infection |
| title_sort | design and production of a novel recombinant chimeric il2 omp31 antigen against brucella infection |
| topic | Brucella melitensis Interleukin Omp31 Cytokines |
| url | http://archrazi.areeo.ac.ir/article_116404_4286f2e4b17f23e95d54e0bb1644ae65.pdf |
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