Abstract
Bet v 1 family proteins of the Fagales order are the major pollen allergens. Fourteen genes of the Bet v 1 family were randomly recombined in vitro by means of DNA shuffling. A resultant low allergenic Tree Pollen Chimera 7 (TPC7) has high immunogenicity against multiple Fagales pollen allergy in vitro and in vivo. Towards development of rice seed-based edible vaccine against multiple Fagales pollen allergy, we generated transgenic rice that accumulate TPC7 in the endosperm. TPC7 high accumulating lines were screened by immunoblot using anti-TPC7 antibody. TPC7 in high accumulating lines was detectable as two visible bands on CBB-stained SDS-PAGE gel. After glycosidase reaction, two bands became a single band, indicating that rice seed-derived TPC7 was glycosylated. In rice endosperm cells, seed storage protein prolamins are deposited in the ER-derived protein body-I (PB-I), and glutelins and globulin are deposited in the storage vacuole PB-II. Since TPC7 was deposited into the novel organelle other than PB-I or PB-II, we termed this as TPC7 body. We will report biochemical properties of TPC7 protein in the transgenic rice and the character of TPC7 body.
