1. Production Of Recombinant N Protein Of Infectious Bronchitis Virus Using The Baculovirus Expression System And Its Assessment As A Diagnostic Antigen
- Author
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E. Bayraktar, Maira Cotton-Caballero, Ozge Aydin, Eda Altan Tarakci, Juergen A. Richt, Utku Y. Cizmecigil, Burhan Çetinkaya, Huseyin Yilmaz, Nuri Turan, Bonto Faburay, Aydın Gürel, and Aysun Yilmaz
- Subjects
0106 biological sciences ,Gene Expression ,Expression ,Infectious bronchitis virus ,01 natural sciences ,Applied Microbiology and Biotechnology ,Biochemistry ,N protein ,law.invention ,law ,Baculovirus ,Antigens, Viral ,Purification ,Phylogeny ,2. Zero hunger ,Recombinant ,medicine.diagnostic_test ,Vaccination ,General Medicine ,Nucleocapsid Proteins ,Recombinant Proteins ,3. Good health ,embryonic structures ,Recombinant DNA ,Coronavirus Infections ,Baculoviridae ,Biotechnology ,Turkeys ,S1 ,animal structures ,medicine.drug_class ,Bioengineering ,Elisa ,Spodoptera ,Biology ,Monoclonal antibody ,Antibodies ,Article ,Virus ,Cell Line ,Affinity chromatography ,Western blot ,Antigen ,010608 biotechnology ,medicine ,Animals ,Coronavirus Nucleocapsid Proteins ,Molecular Biology ,Poultry Diseases ,Antiserum ,010405 organic chemistry ,Virology ,0104 chemical sciences ,Chickens - Abstract
The avian coronavirus-infectious bronchitis virus (AvCoV-IBV) is recognized as an important avian pathogen, and new viral variants are a continuous threat to the poultry industry worldwide. Sensitive diagnostics and efficacious vaccines are necessary to combat IBV infections in chickens. The aim of this study was to produce recombinant N protein of IBV in the baculovirus system to use in ELISA diagnostic tests in order to enable the assessment of the sero-prevalence and risk of IBV infections in chickens in Turkey. For this, the gene encoding the N protein of the Beaudette strain of IBV was expressed using a recombinant baculovirus expression system. The recombinant N protein was purified using Ni-NTA affinity chromatography. An estimated 50-kDa recombinant protein corresponding to the expected molecular weight of IBV N including the 6xHis tag was detected using an anti-His monoclonal antibody. Specific immunoreactivity of the recombinant protein was confirmed by Western blot using antiserum obtained from vaccinated and naturally infected chicken from Turkey as well as using a monoclonal antibody raised against the N protein of the IBV Massachusetts strain. The results obtained with the in-house ELISA had high agreement with a commercial ELISA. Immunoreactivity analysis using antisera in Western blotting and the in-house ELISA suggests that the recombinant IBV N protein could be broadly cross-reactive with antisera produced against different IBV strains. We conclude that the recombinant baculovirus expressed IBV N protein could serve as a useful diagnostic antigen for detection of IBV infections in chickens by ELISA. TUBITAK: Scientific and Technological Research Council of TurkeyTurkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [113O411]; Developing Scholars Program (DSP), Kansas State University, Office of Undergraduate Research and Creative Inquiry; DHS Center of Excellence for Emerging and Zoonotic Animal Diseases (CEEZAD) This study was funded by TUBITAK: Scientific and Technological Research Council of Turkey (Project No: 113O411), Developing Scholars Program (DSP), Kansas State University, Office of Undergraduate Research and Creative Inquiry; DHS Center of Excellence for Emerging and Zoonotic Animal Diseases (CEEZAD).
- Published
- 2019