In planta expression of specific single chain fragment antibody (scFv) against nucleocapsid protein of fig mosaic virus (FMV)

IF 2.2 4区 医学 Q3 BIOCHEMICAL RESEARCH METHODS Journal of virological methods Pub Date : 2024-02-17 DOI:10.1016/j.jviromet.2024.114904
Mahsa Jahandideh , Farshad Rakhshandehroo , Mohammad Reza Safarnejad , Amir Sahraroo , Toufic Elbeaino
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Abstract

Fig mosaic virus (FMV) is recognized as the main viral agent associated with the mosaic disease (MD) of fig trees (Ficus carica). Due to its worldwide occurrence, FMV represents the most significant global threat to the production of fig fruit. A disease management strategy against the MD in fig orchards has never been effective; and therefore, expression of recombinant antibody in plant cells could provide an alternative approach to suppress FMV infections. In this study we focused on expressing a specific recombinant antibody, a single-chain variable fragment (scFv), targeting the nucleocapsid protein (NP) of FMV in planta. To accomplish this objective, we inserted the scFv gene into a plant expression vector and conducted transient expression in leaves of Nicotiana tabacum cv. Samson plants. The construct was transiently expressed in tobacco plants by agroinfiltration, and antibody of the anticipated size was detected by immunoblotting. The produced plantibody was then assessed for specificity using ELISA and confirmed by Western blot analysis. In this study, the plantibody developed against FMV could be considered as a potential countermeasure to the infection by conferring resistance to MD.

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在植物体内表达针对无花果花叶病毒(FMV)核壳蛋白的特异性单链片段抗体(scFv)。
无花果马赛克病毒(FMV)被认为是与无花果树(Ficus carica)马赛克病(MD)相关的主要病毒病原体。无花果花叶病毒在世界各地都有发生,是全球无花果生产面临的最大威胁。在无花果园中,针对无花果马赛克病的病害管理策略从未奏效;因此,在植物细胞中表达重组抗体可为抑制 FMV 感染提供另一种方法。本研究的重点是在植物体内表达一种特异性重组抗体--单链可变片段(scFv),靶向 FMV 的核壳蛋白(NP)。为实现这一目标,我们将 scFv 基因插入植物表达载体,并在烟草植物 Samson 的叶片中进行瞬时表达。通过农业渗透法在烟草植物中瞬时表达构建体,并通过免疫印迹法检测预期大小的抗体。然后用酶联免疫吸附法评估所产生的植物抗体的特异性,并通过 Western 印迹分析加以确认。在这项研究中,针对 FMV 开发的植物抗体可通过赋予 MD 抗性而被视为一种潜在的抗感染措施。
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来源期刊
CiteScore
5.80
自引率
0.00%
发文量
209
审稿时长
41 days
期刊介绍: The Journal of Virological Methods focuses on original, high quality research papers that describe novel and comprehensively tested methods which enhance human, animal, plant, bacterial or environmental virology and prions research and discovery. The methods may include, but not limited to, the study of: Viral components and morphology- Virus isolation, propagation and development of viral vectors- Viral pathogenesis, oncogenesis, vaccines and antivirals- Virus replication, host-pathogen interactions and responses- Virus transmission, prevention, control and treatment- Viral metagenomics and virome- Virus ecology, adaption and evolution- Applied virology such as nanotechnology- Viral diagnosis with novelty and comprehensive evaluation. We seek articles, systematic reviews, meta-analyses and laboratory protocols that include comprehensive technical details with statistical confirmations that provide validations against current best practice, international standards or quality assurance programs and which advance knowledge in virology leading to improved medical, veterinary or agricultural practices and management.
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