بیان ژن کد کننده آنتی ژن VP1 و بررسی امکان زراعت مولکولی آن جهت تولید واکسن نوترکیب تب برفکی در گیاه توتون (Nicotiana tabacum)

Foot and Mouth Disease (FMD) is a serious livestock disease bearing severe economical damage on milk and meet production industries. Currently, attenuated viruses are used as vaccine against the disease. However, high production cost and the risk of activation of attenuated virus in traditional vaccine systems have attracted many investigators to production of recombinant vaccines against FMD. In the present study, an epitope-based approach was implemented for production of FMD recombinant vaccine in tobacco plant (Nicotiana tabacum). To achieve this, a DNA sequence encoding 129-169 amino acids of VP1 capsid protein was expressed in tobacco via agrobacterium-mediated genetic transformation. Putative transgenic lines were selected by PCR and kanamycin resistance and transgene expression in these lines was evaluated in both transcription and translation levels. Although all transgenic lines showed positive signals in Real Time PCR and ELISA assays, the recombinant protein band in western blotting was only present in two lines namely 10 and 42. Efficiency of the recombinant protein to induce immune response in model animal (rabbit) was further investigated. As confirmed by ELISA, it was revealed that the recombinant protein was able to induce specific antibodies in immunized animals. Moreover, since a primary goal of the investigation was to enhance expression of the transgene by adopting strategies such as application of ribosome binding site, signal peptide and codon optimization, expression of the recombinant protein was quantified in lines 10 and 42. It was observed that transgene expression level in lines 10 and 42 was 0.65% and 0.72% of total soluble proteins. In summary, the results obtained in the present investigation indicated that an efficient recombinant vaccine against FMD can be achieved by implementing appropriate epitopes and suitable strategies.