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MOLECULAR CLONING AND FUNCTIONAL ANALYSIS OF PFWD, A PUTATIVE REGULATORY PROTEIN, IN ANTHOCYANIN BIOSYNTHESIS FROM PERILLA FRUTESCENS 

作成者 Sompornpailin, Kanokporn
キーワード等 anthocyanin biosyn thesis, Perilla frutescens WD repeats
日本十進分類法 (NDC) 499
内容 学位:千大院薬博甲第薬134号
抄録:The WD-repeat proteins are found in eukaryotes and play an important role in the regulation of a wide variety of cellular functions such as signal transduction, transcription, and proliferation. In this study, I investigated the cloning of a novel WD40-repeat gene, Pfwd, from Perilla frutescens by screening cDNA library of the red leaves P. frutescens with anll cDNA as the probe from Petunia hybrida, which encodes a regulatory protein of anthocyanin biosynthesis. The characterization of full-length cDNA revealed that Pfwd cDNA contains 1411 base pairs, encoding a predicted protein of 333 amino acids. The deduced amino acid sequence shows 81.3 0dentity with that of an 11 from P. hybrida. The C-terminal region contains WD repeat that is highly conserved in several proteins even from the organisms such as yeast, nematodes and mammals that do not produce anthocyanin. Numerous sites of putative post-translational modification, such as N-linked glycosylation, myristoylation and phosphorylation sites, were also identified. Overexpression of PFWD in Arabidopsis and Escherichia coli, subcellular localization of PFWD and protein-protein interaction of PFWD and other regulatory factors were investigated to examine the role of PFWD in the regulation of anthocyanin biosynthesis. The transgenic Arabidopsis plants overexpressing PFWD exhibited various phenotypic changes including enhancement of anthocyanin production. Overexpression of PFWD affected to viability of plant. Overexpression in E. coli showed weak expression band of maltose binding protein (MBP) and PFWD fusion. The fusion protein of PFWD and β-glucuronidase (GUS) transiently expressed in onion epidermal cells was localized in the cytosol under both dark and light conditions. The study of interaction among PFWD and anthocyanin regulatory proteins in yeast two-hybrid system showed the strong interaction between PFWD and MYC-RP, and between MYB-P1 and MYC-RP.…

コンテンツの種類 博士論文 Doctoral Thesis
DCMI資源タイプ text
ファイル形式 application/pdf
ハンドルURL http://mitizane.ll.chiba-u.jp/meta-bin/mt-pdetail.cgi?cd=00022535
フルテキストへのリンク http://mitizane.ll.chiba-u.jp/metadb/up/assist1/Y_K-013.pdf
言語 英語


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Perilla frutescens WD repeats