{"title":"盒式构建及在酿酒酵母中共同表达甲型 H1N1 流感病毒和新城疫病毒的 NA 和 F 蛋白","authors":"A. Seyed Hamzehzadeh, A. Hesampour","doi":"10.3103/s0891416823040134","DOIUrl":null,"url":null,"abstract":"<p>The aim of this study is to optimize the design, construction, and expression of recombinant protein, NA-F, derived from pathogenic influenza and Newcastle viruses within the eukaryotic host <i>Saccharomyces cerevisiae</i>. The chimeric gene was constructed using a homologous recombination method, and the target genes were cloned onto the T&A vector using PCR cloning techniques. Subsequently, the chimeric gene was expressed in <i>S. cerevisiae</i>, and its presence was confirmed through extracellular expression validation, Western blotting, and specific antibody detection. The recombinant protein displayed significant biological activity, demonstrating its functional characteristics and potential for use in immunization. The chimeric gene structure produced in this study holds promise as a candidate to replace existing vaccines worldwide, considering the high prevalence of influenza and Newcastle viruses in poultry and the need for a universal subunit vaccine. However, further research in animal and human models is essential to ensure safety before widespread application.</p>","PeriodicalId":0,"journal":{"name":"","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Cassette Construction and Co-Expression of NA and F Proteins of H1N1 Influenza Virus and Newcastle Disease Virus in Saccharomyces cerevisiae\",\"authors\":\"A. Seyed Hamzehzadeh, A. Hesampour\",\"doi\":\"10.3103/s0891416823040134\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>The aim of this study is to optimize the design, construction, and expression of recombinant protein, NA-F, derived from pathogenic influenza and Newcastle viruses within the eukaryotic host <i>Saccharomyces cerevisiae</i>. The chimeric gene was constructed using a homologous recombination method, and the target genes were cloned onto the T&A vector using PCR cloning techniques. Subsequently, the chimeric gene was expressed in <i>S. cerevisiae</i>, and its presence was confirmed through extracellular expression validation, Western blotting, and specific antibody detection. The recombinant protein displayed significant biological activity, demonstrating its functional characteristics and potential for use in immunization. The chimeric gene structure produced in this study holds promise as a candidate to replace existing vaccines worldwide, considering the high prevalence of influenza and Newcastle viruses in poultry and the need for a universal subunit vaccine. However, further research in animal and human models is essential to ensure safety before widespread application.</p>\",\"PeriodicalId\":0,\"journal\":{\"name\":\"\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0,\"publicationDate\":\"2024-03-20\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://doi.org/10.3103/s0891416823040134\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.3103/s0891416823040134","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
摘要
本研究旨在优化重组蛋白NA-F的设计、构建和表达,NA-F来源于真核宿主酿酒酵母(Saccharomyces cerevisiae)中的致病性流感病毒和新城疫病毒。采用同源重组方法构建了嵌合基因,并利用 PCR 克隆技术将目标基因克隆到 T&A 载体上。随后,嵌合基因在 S. cerevisiae 中表达,并通过胞外表达验证、Western 印迹和特异性抗体检测证实了其存在。重组蛋白显示出显著的生物活性,证明了其功能特性和用于免疫的潜力。考虑到流感和新城疫病毒在家禽中的高流行率以及对通用亚单位疫苗的需求,本研究中产生的嵌合基因结构有望成为替代全球现有疫苗的候选产品。不过,在广泛应用之前,必须在动物和人体模型中开展进一步研究,以确保安全性。
Cassette Construction and Co-Expression of NA and F Proteins of H1N1 Influenza Virus and Newcastle Disease Virus in Saccharomyces cerevisiae
The aim of this study is to optimize the design, construction, and expression of recombinant protein, NA-F, derived from pathogenic influenza and Newcastle viruses within the eukaryotic host Saccharomyces cerevisiae. The chimeric gene was constructed using a homologous recombination method, and the target genes were cloned onto the T&A vector using PCR cloning techniques. Subsequently, the chimeric gene was expressed in S. cerevisiae, and its presence was confirmed through extracellular expression validation, Western blotting, and specific antibody detection. The recombinant protein displayed significant biological activity, demonstrating its functional characteristics and potential for use in immunization. The chimeric gene structure produced in this study holds promise as a candidate to replace existing vaccines worldwide, considering the high prevalence of influenza and Newcastle viruses in poultry and the need for a universal subunit vaccine. However, further research in animal and human models is essential to ensure safety before widespread application.