Cassette Construction and Co-Expression of NA and F Proteins of H1N1 Influenza Virus and Newcastle Disease Virus in Saccharomyces cerevisiae

IF 0.4 4区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Molecular Genetics, Microbiology and Virology Pub Date : 2024-03-20 DOI:10.3103/s0891416823040134

A. Seyed Hamzehzadeh, A. Hesampour

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Abstract

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.

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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

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。

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来源期刊

Molecular Genetics, Microbiology and Virology BIOCHEMISTRY & MOLECULAR BIOLOGY-MICROBIOLOGY

CiteScore

0.70

自引率

0.00%

发文量

审稿时长

>12 weeks

期刊介绍： Molecular Genetics, Microbiology and Virology is a journal that covers most topical theoretical and applied problems of molecular genetics of pro- and eukaryotic organisms, molecular microbiology and molecular virology. An important part the journal assigns to investigations of the genetic apparatus of microorganisms, searching for forms of genetic exchange, genetic mapping of pathogenic causative agents, to ascertainment of the structure and functions of extrachromosomal factors of heredity and migratory genetic elements, to theoretical studies into the mechanisms of genetic regulation. The journal publishes results of research on molecular and genetic bases of an eukaryotic cell, functioning of chromosomes and chromatin, nature of genetic changes in malignization and a set of hereditary diseases. On the pages of the journal there is covered the formulation of molecular bases of virology including issues of integration of viral and cellular genomes, and issues of persistence. The journal plans to put materials on genetic engineering, envisaging synthesis and isolation of genes from natural reservoirs, creation of plasmid- and virus-based vector, production of recombinant DNA molecules, the creation of Gene Banks for Microbes, animals, and human; and also on biotechnological production of hormones, components of antiviral vaccines, diagnostic and therapeutic preparations.