| Products/Services Used | Details | Operation |
|---|---|---|
| Peptide Synthesis | The DNA sequences encoding Mal d 1, Bet v 1 and TPC7 were optimized for translation based on the codons frequently used in rice seed storage protein gens and their DNA sequences were synthesized by GenScript Corporation (NJ, USA). The N terminal signal peptide of the glutelin GluB-1 that is required for translocation into the ER and the KDEL ER retention signals were attached to their N and C termini, respectively. The gene encoding the green fluorescence protein (GFP) reporter was inserted in frame between the GluB-1 signal and these PR10 related genes. In order to express specifically in the endosperm or constitutively in vegetative tissues, 2.3kb glutelin GluB-1 promoter or maize ubiquitin promoter were ligated upstream of these three fusion constructs, which were followed by the 0.65kb of glutelin GluB-1terminator (Fig. 1B). | Get A Quote |
Apple food allergen Mal d 1 and birch pollen allergen Bet v 1 belong to the same pathogen related protein 10 (PR10) family. When each of these allergens was expressed as a secretory protein by fusion with the GFP reporter in transgenic rice by ligating an N terminal signal peptide and a C terminal KDEL ER retention signal under the control of the maize ubiquitin constitutive promoter, the GFP:Mald1 highly accumulated in various tissues, whereas the accumulation level of GFP:Betv1 was remarkably reduced in vegetative tissues except for seed. Analysis by RT-PCR exhibited that there was little difference in transcript levels between them, indicating the involvement of post-transcriptional regulation. To investigat... More
