Maziar Habibi; Saeid Malekzadeh; hasan marashi; Afsaneh Mohkami
Abstract
Regarding high potential of green plants for development of recombinant vaccines, this research was conducted to evaluate expression of a novel recombinant vaccines against Foot and Mouth Disease (FMDV) in tobacco plant. For this purpose, a synthetic gene encoding 129-169 amino acids of foot and mouth ...
Read More
Regarding high potential of green plants for development of recombinant vaccines, this research was conducted to evaluate expression of a novel recombinant vaccines against Foot and Mouth Disease (FMDV) in tobacco plant. For this purpose, a synthetic gene encoding 129-169 amino acids of foot and mouth disease virus capsid protein VP1 was transferred to tobacco plant via Agrobacterium-mediated genetic transformation. Considering codon usage optimized for tobacco, ribosome binding site and endoplasmic reticulum signal peptide were included in the synthetic gene to enhance expression level. Expression of the synthetic gene in tobacco seedlings was analyzed at transcription and translation levels and production of recombinant protein was quantified. Moreover, an in vivo immunization assay was carried out to verify immunogenicity of the expressed peptide in model animals. Results showed that expression of the recombinant protein in two lines of transgenic plants was a high as 0.65% and 0.72% of total soluble protein. The recombinant protein was able to induce immunogenic response when parenterally administered in rabbit.
Molecular biology and (trans) gene expression technology in plant breeding
Maziar Habibi; Saeid Malekzadeh-Shafaroudi; Hassan Marashi; Nasrin Moshtaghi; Mohammadreza Nasiri; Saeed Zibaee
Abstract
Foot and Mouth Disease (FMD) is a very dangerous livestock disease which causes a serious loss in the production of milk and meat. Therefore, producing an effective recombinant subunit vaccine virus this disease is of great importance. Transient gene expression is a valuable tool to reach rapid and acceptable ...
Read More
Foot and Mouth Disease (FMD) is a very dangerous livestock disease which causes a serious loss in the production of milk and meat. Therefore, producing an effective recombinant subunit vaccine virus this disease is of great importance. Transient gene expression is a valuable tool to reach rapid and acceptable recombinant vaccine. An Agrobacterium-mediated transient gene expression assay was carried out in spinach (Spinacia oleracea) 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 foreign gene and its expression in transformed plants were confirmed through polymerase chain reaction (PCR), real time PCR, protein Dot blot and ELISA. The results obtained in this examination showed quite a high level of gene expression in spinach 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.