Engineering and Development of VP4-VP7 Rotavirus Fusion Protein in Pichia pastoris
Introduction: Vaccination is the most effective measure to prevent Rotavirus infection in children under five years of age. The most important targets of neutralizing and protective antibodies against this virus are VP4 and VP7 proteins of Rotavirus. Today, the recombinant protein produced in yeast...
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| Format: | Article |
| Language: | English |
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Pasteur Institute of Iran
2020-12-01
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| Series: | Vaccine Research |
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| Online Access: | http://vacres.pasteur.ac.ir/article-1-263-en.pdf |
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| author | Mostafa Amopour Bahnamiry Hooman Kaghazian Seyed Dawood Mousavi-Nasab Mohammad Reza Aghasadeghi Golnaz Bahramali |
| author_facet | Mostafa Amopour Bahnamiry Hooman Kaghazian Seyed Dawood Mousavi-Nasab Mohammad Reza Aghasadeghi Golnaz Bahramali |
| author_sort | Mostafa Amopour Bahnamiry |
| collection | DOAJ |
| description | Introduction: Vaccination is the most effective measure to prevent Rotavirus infection in children under five years of age. The most important targets of neutralizing and protective antibodies against this virus are VP4 and VP7 proteins of Rotavirus. Today, the recombinant protein produced in yeast plays an important role in producing highly effective and cost-effective vaccines. Methods: The effect of different linkers, including flexible and rigid, were evaluated on the stability and immunogenicity of the protein via in silico assays. A suitable linker was selected and expressed in Pichia pastoris yeast. Prediction and validation were carried out using bioinformatics tools, including Expasy's ProtPara, Phyre2 online server, I-TASSER server, ElliPro. Moreover, an appropriate linker was selected for cloning into pPICZα and expression in P. pastoris. Results: The results showed that as a flexible linker, (GGGGS)3 was the best structure to provide stability for VP4-VP7 target fusion protein. The produced recombinant protein was stable after the expression. Conclusion: These in silico results and expression data on P. pastoris suggested that VP4–(GGGGS)3-VP7 construct can potentially serve as a potent immunogenic candidate for recombinant Rotavirus vaccines. |
| first_indexed | 2024-12-10T10:33:10Z |
| format | Article |
| id | doaj.art-15ff5b1d85684d6abed053a1353a4eea |
| institution | Directory Open Access Journal |
| issn | 2383-2819 2423-4923 |
| language | English |
| last_indexed | 2024-12-10T10:33:10Z |
| publishDate | 2020-12-01 |
| publisher | Pasteur Institute of Iran |
| record_format | Article |
| series | Vaccine Research |
| spelling | doaj.art-15ff5b1d85684d6abed053a1353a4eea2022-12-22T01:52:30ZengPasteur Institute of IranVaccine Research2383-28192423-49232020-12-017210110610.52547/vacres.7.2.101Engineering and Development of VP4-VP7 Rotavirus Fusion Protein in Pichia pastorisMostafa Amopour Bahnamiry0https://orcid.org/0000-0002-0542-4000Hooman Kaghazian1https://orcid.org/0000-0003-4301-3381Seyed Dawood Mousavi-Nasab2https://orcid.org/0000-0002-4831-3108Mohammad Reza Aghasadeghi3https://orcid.org/0000-0003-2186-0774Golnaz Bahramali https://orcid.org/0000-0002-1762-4907Department of Research and Development, Production and Research Complex, Pasteur Institute of Iran, Tehran, IranDepartment of Research and Development, Production and Research Complex, Pasteur Institute of Iran, Tehran, IranViral Vaccine Research Center, Pasteur Institute of Iran, Tehran, Iran Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran. Introduction: Vaccination is the most effective measure to prevent Rotavirus infection in children under five years of age. The most important targets of neutralizing and protective antibodies against this virus are VP4 and VP7 proteins of Rotavirus. Today, the recombinant protein produced in yeast plays an important role in producing highly effective and cost-effective vaccines. Methods: The effect of different linkers, including flexible and rigid, were evaluated on the stability and immunogenicity of the protein via in silico assays. A suitable linker was selected and expressed in Pichia pastoris yeast. Prediction and validation were carried out using bioinformatics tools, including Expasy's ProtPara, Phyre2 online server, I-TASSER server, ElliPro. Moreover, an appropriate linker was selected for cloning into pPICZα and expression in P. pastoris. Results: The results showed that as a flexible linker, (GGGGS)3 was the best structure to provide stability for VP4-VP7 target fusion protein. The produced recombinant protein was stable after the expression. Conclusion: These in silico results and expression data on P. pastoris suggested that VP4–(GGGGS)3-VP7 construct can potentially serve as a potent immunogenic candidate for recombinant Rotavirus vaccines.http://vacres.pasteur.ac.ir/article-1-263-en.pdfrotavirusvaccinebioinformaticppiczαpichia pastoris |
| spellingShingle | Mostafa Amopour Bahnamiry Hooman Kaghazian Seyed Dawood Mousavi-Nasab Mohammad Reza Aghasadeghi Golnaz Bahramali Engineering and Development of VP4-VP7 Rotavirus Fusion Protein in Pichia pastoris Vaccine Research rotavirus vaccine bioinformatic ppiczα pichia pastoris |
| title | Engineering and Development of VP4-VP7 Rotavirus Fusion Protein in Pichia pastoris |
| title_full | Engineering and Development of VP4-VP7 Rotavirus Fusion Protein in Pichia pastoris |
| title_fullStr | Engineering and Development of VP4-VP7 Rotavirus Fusion Protein in Pichia pastoris |
| title_full_unstemmed | Engineering and Development of VP4-VP7 Rotavirus Fusion Protein in Pichia pastoris |
| title_short | Engineering and Development of VP4-VP7 Rotavirus Fusion Protein in Pichia pastoris |
| title_sort | engineering and development of vp4 vp7 rotavirus fusion protein in pichia pastoris |
| topic | rotavirus vaccine bioinformatic ppiczα pichia pastoris |
| url | http://vacres.pasteur.ac.ir/article-1-263-en.pdf |
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