Hernández-Aviña Ana Del Socorro, Mendoza-Gómez Luis Fernando, Hernández-Mancillas Xitlalli D, Salazar-González Jorge Alberto, Zapata-Cuellar Lorena, Camacho-Ruiz Rosa María, Comas-García Mauricio, Sarmiento-Silva Rosa Elena, Rodríguez Jorge A, Arellano-Plaza Melchor, Flores-Valdez Mario Alberto, Gutiérrez-Ortega Abel
Unidad de Biotecnología Médica y Farmacéutica, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco, Normalistas 800, Colinas de La Normal, Jalisco, 44270, México.
Unidad de Biotecnología Industrial, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco, Camino Arenero 1227, El Bajío del Arenal, Jalisco, 45019, México.
Mol Biotechnol. 2024 Nov 30. doi: 10.1007/s12033-024-01334-z.
Porcine circovirus type 2 (PCV2) is a pervasive pathogen in the swine industry, leading to a spectrum of disorders known as porcine circovirus associated diseases (PCVAD). The PCV2 Cap protein contains critical antigenic epitopes and is the primary target for vaccine development. Current vaccines include inactivated viral particles and virus-like particles (VLPs), with experimental vaccines exploring various innovative approaches. This study introduces a novel Golden Gate assembly-based, Escherichia coli expression destination plasmid, pPRSVCP-E18, designed for the in-frame insertion of short peptide-coding DNA sequences into a highly antigenic site of Papaya ringspot virus (PRSV) coat protein (CP). We successfully cloned four PCV2 Cap peptides into this plasmid, expressed the chimeric CP proteins in E. coli under flask and bioreactor conditions, and assembled them into VLPs. These VLPs, when adjuvated and administered to BALB/c mice, elicited a specific IgG response against the PCV2 Cap peptides. Our findings demonstrate the plasmid's high efficiency for Golden Gate cloning and its potential in developing subunit vaccines against PCV2, contributing to the sustainable control of PCVAD in the swine industry.
猪圆环病毒2型（PCV2）是养猪业中一种普遍存在的病原体，可导致一系列被称为猪圆环病毒相关疾病（PCVAD）的病症。PCV2衣壳蛋白包含关键的抗原表位，是疫苗开发的主要靶点。目前的疫苗包括灭活病毒颗粒和病毒样颗粒（VLP），实验性疫苗正在探索各种创新方法。本研究介绍了一种基于金门组装的新型大肠杆菌表达目的质粒pPRSVCP-E18，其设计用于将短肽编码DNA序列读框内插入番木瓜环斑病毒（PRSV）外壳蛋白（CP）的高抗原性位点。我们成功地将四种PCV2衣壳肽克隆到该质粒中，在摇瓶和生物反应器条件下在大肠杆菌中表达嵌合CP蛋白，并将它们组装成VLP。这些VLP在佐剂化并给予BALB/c小鼠后，引发了针对PCV2衣壳肽的特异性IgG反应。我们的研究结果证明了该质粒在金门克隆中的高效性及其在开发针对PCV2的亚单位疫苗方面的潜力，有助于养猪业中PCVAD的可持续防控。
