Development of transgenic tobacco and potato plants for expression of mite allergens Der p II and Der pV

• Main Authors: Hsiao-Li Wang, 王曉俐
• Other Authors: Hsin-Mei Ku
• Format: Others
• Language: zh-TW
• Published: 2003
• Online Access: http://ndltd.ncl.edu.tw/handle/84301918073488426798

碩士 === 國立中興大學 === 農藝學系 === 91 === Allergic diseases have been increasing worldwide and causing problems for millions of children. Dust mite allergens are the most common cause of perennial allergic rhinitis and asthma . Edible allergen vaccine is a new form of treatment to reduce allergic response to allergens. The purified allergen vaccine is a very expensive treatment for allergic diseases. The focus of our research was to develop transgenic plants for expressing mite allergen as an edible and low cost vaccine for oral tolerance to allergy. We have generated plant expression and transformation vectors containing mite allergen genes Der p II and Der p V. The mite allergen genes were placed under control of 35S promoter of Cauliflower mosaic virus and nopaline synthase terminator of Agrobacterium tumefaciens. At the 3´-end of Der p II and Der p V, a nucleotide sequence coding for the hexapeptide Ser-Gln-Lys-Asn-Gln-Leu (SEKDEL) was added for retention of the protein in the endoplasmic reticulum (ER). The Der p II and Der p V genes were introduced into tobacco through the antigen gene-harboring A. tumefaciens-mediated plant transformation method as a model system. Expression of the introduced genes were confirmed by PCR, RT-PCR and Western blot analysis, which indicated that transformed plants expressed Der p II and Der p V and displayed structural and immunological characteristics identical to the Escherichia coli expressed protein. Our aim was to express mite antigens in edible plant, therefore, the same constructions was used to generate transgenic potato expressing Der p II and Der p V. Integration of the transgene in potato was also confirmed by PCR, RT-PCR and Southern blot analysis and expression of the Der p II and Der p V protein was confirmed by Western blot analysis. The transgeinc tobacco and potato plants expressing the transgene carrying the SEKDEL retention signal showed higher expression levels of Der p II and Der p V allergic proteins. The maximum level that could be detected was 0.01% of the total soluble proteins. This report first presented the expression of mite allergens in plant tissues with an immunogenic form and the transgenic plants generated by this method may provide a model system for development of low-cost edilbe oral vaccine.