Author/Authors :
Habibi-Pirkoohi، Maziar نويسنده Department of Biotechnology and Plant Breeding, Ferdowsi University of Mashhad, Mashhad, Iran , , Malekzadeh-Shafaroudi، Saeid نويسنده Department of Biotechnology and plant breeding,Ferdowsi University of Mashhad,Mashhad,Iran , , Marashi، Hasan نويسنده Department of Biotechnology and plant breeding,Ferdowsi University of Mashhad,Mashhad,Iran , , Zibaee، Saeid نويسنده Razi Vaccine and Serum Research Institute,Ministry of Agriculture,Mashhad,Iran , , Mohkami، Afsaneh نويسنده Research and Technology Institute of Plant production,Shahid Bahonar University,Kerman,Iran , , Nejatizadeh، Saba نويسنده Department of Biotechnology and plant breeding,Ferdowsi University of Mashhad,Mashhad,Iran ,
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.
Keywords :
FMDV , agroinfiltration , Recombinant vaccine , alfalfa , VP1
