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.

本研究的目的是优化源自真核宿主酿酒酵母内致病性流感和新城疫病毒的重组蛋白 NA-F 的设计、构建和表达。采用同源重组方法构建嵌合基因，并采用PCR克隆技术将目的基因克隆到T&A载体上。随后，该嵌合基因在酿酒酵母中表达，并通过胞外表达验证、Western blotting和特异性抗体检测证实其存在。重组蛋白表现出显着的生物活性，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疫苗。然而，在动物和人体模型中进一步研究对于确保广泛应用之前的安全性至关重要。