Transient Expression of Coat Protein of Foot and Mouth Disease Virus (FMDV) in Alfalfa (Medicago sativa) by Agroinfiltration

M. Habibi-Pirkoohi, S. Malekzadeh-Shafaroudi, H. Marashi, S. Zibaee, A. Mohkami, Saba NejatizaXeh
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引用次数: 2

Abstract

An Agrobacterium-mediated transient gene expression assay was carried out in alfalfa (Medicago sativa) leaves for expression of a chimeric gene encoding a part of capsid protein of Foot and Mouth Disease virus called VP1. The plant leaves were transformed via agroinfiltration procedure. The presence of the foreign gene and its expression in transformed plants were evaluated by polymerase chain reaction (PCR), real time PCR, protein Dot blot and ELISA. Moreover, gene expression in the transformed leaves was quantified by ELISA method. The results obtained in this investigation indicated high level of gene expression in alfalfa leaves, showing that transient gene expression can be applied as an effective and time-saving procedure for the production of recombinant proteins. The procedures for transformation, detection of recombinant protein and its application for molecular experiments are described in the study.
苜蓿口蹄疫病毒(FMDV)外壳蛋白在农业渗透中的瞬时表达
利用农杆菌介导的瞬时基因表达试验,在紫花苜蓿叶片中表达了一个编码口蹄疫病毒衣壳蛋白部分的嵌合基因VP1。植物叶片通过农业渗透转化。采用聚合酶链反应(PCR)、实时荧光定量PCR、蛋白斑点杂交(protein Dot blot)和酶联免疫吸附法(ELISA)检测外源基因的存在及其在转化植株中的表达。利用酶联免疫吸附法(ELISA)对转化叶片中的基因表达量进行定量分析。本研究结果表明,该基因在紫花苜蓿叶片中有较高的表达水平,表明瞬时基因表达可以作为一种有效且省时的重组蛋白生产方法。介绍了重组蛋白的转化、检测及其在分子实验中的应用。
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