Journals / Turkish Journal of Veterinary and Animal Sciences / 2011 / Cilt: 35 - Sayı: 4
Expression and detection of the FMDV VP1 transgene and expressed structural protein in Arabidopsis thaliana
- Pages
- 235–242
- DOI
- —
Abstract
To explore the feasibility of developing a new type of plantderived foot-and-mouth disease virus (FMDV) oral vaccine, the plant seed-specific expression vector p7SBin438/VP1 carrying the VP1 gene of the FMDV strain O/China/99 was constructed and transformed into Agrobacterium tumefaciens strain GV3101. This strain was used for transformation of Arabidopsis thaliana via the floral-dip method. The kanamycin-resistant transgenic plants were selected, and the VP1 gene and protein expressions were confirmed by PCR and sandwich-ELISA, respectively. The ELISA positive plants were further confirmed by Western blot assay, and the VP1 gene in the transgenic progeny was confirmed by PCR. The results showed that the Agrobacterium-mediated floral-dip transformation method could produce transgenic Arabidopsis containing the VP1 gene. Therefore, this protocol provides a successful strategy to deliver viral genes to leguminous plants for genetic studies and for production of low-cost oral vaccines.
Özet
To explore the feasibility of developing a new type of plantderived foot-and-mouth disease virus (FMDV) oral vaccine, the plant seed-specific expression vector p7SBin438/VP1 carrying the VP1 gene of the FMDV strain O/China/99 was constructed and transformed into Agrobacterium tumefaciens strain GV3101. This strain was used for transformation of Arabidopsis thaliana via the floral-dip method. The kanamycin-resistant transgenic plants were selected, and the VP1 gene and protein expressions were confirmed by PCR and sandwich-ELISA, respectively. The ELISA positive plants were further confirmed by Western blot assay, and the VP1 gene in the transgenic progeny was confirmed by PCR. The results showed that the Agrobacterium-mediated floral-dip transformation method could produce transgenic Arabidopsis containing the VP1 gene. Therefore, this protocol provides a successful strategy to deliver viral genes to leguminous plants for genetic studies and for production of low-cost oral vaccines.