Control of foreign polypeptide localization in specific layers of protein body type I in rice seed

Plant Cell Reports
Ai SasouTakehiro Masumura

Abstract

Prolamin-GFP fusion proteins, expressed under the control of native prolamin promoters, were localized in specific layers of PB-Is. Prolamin-GFP fusion proteins were gradually digested from outside by pepsin digestion. In rice seed endosperm, protein body type I (PB-I) has a layered structure consisting of prolamin species and is the resistant to digestive juices in the intestinal tract. We propose the utilization of PB-Is as an oral vaccine carrier to induce mucosal immune response effectively. If vaccine antigens are localized in a specific layer within PB-Is, they could be protected from gastric juice and be delivered intact to the small intestine. We observed the localization of GFP fluorescence in transgenic rice endosperm expressing prolamin-GFP fusion proteins with native prolamin promoters, and we confirmed that the foreign proteins were located in specific layers of PB-Is artificially. Each prolamin-GFP fusion protein was localized in specific layers of PB-Is, such as the outer-most layer, middle layer, and core region. Furthermore, to investigate the resistance of prolamin-GFP fusion proteins against pepsin digestion, we performed in vitro pepsin treatment. Prolamin-GFP fusion proteins were gradually digested from the...Continue Reading

References

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Oct 8, 2008·Bioscience, Biotechnology, and Biochemistry·Yuhi SaitoTakehiro Masumura
Jan 9, 2009·Journal of Experimental Botany·Yuhi SaitoTakehiro Masumura
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Methods Mentioned

BETA
transgenic
PCR
Protein Assay
transmission electron microscopy
fluorescence microscopy

Software Mentioned

Kodak 1D Image Analysis

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