Production of Full-Length Pea Phytochrome A (Type I) Apoprotein by Yeast Expression System

概要

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Pea phytochrome A (type I phytochrome) cDNA spanning the complete coding region was expressed using a yeast expression system. Anti-pea phytochrome A (phyA) antibodies that recognize different epitopes located from the N- to the C-terminus of the molecule detected a single product with a molecular weight of 120 kDa as native pea phyA in western blot analysis of the yeast lysate. This indicated that the yeast system produced full-length pea phyA apoprotein (PHYA). The yield of PHYA was 0.037% of total yeast protein. When size exclusion column chromatography was performed, PHYA expressed in yeast was detected at around 450 kDa although pea phyA was detected at 400 kDa, suggesting that PHYA exists in a nonglobular dimeric form but its conformation is slightly different than that of native pea phyA.
日本植物生理学会の論文