46 タンパク局在解析への改変緑色蛍光タンパクの利用

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Green fluorescent protein has been used as the reporter to examine protein localization in living cell, and there are two variants (CFP and YFP) that have different fluorescence emission spectra. We tested the usefulness of CFP and YFP for two-color labeling of organelle or for protein localization analysis in plant. The result demonstrated that combination of CFP and YFP was suitable for these observations and that use of two types of constructs was sufficient for transient co-expression by particle bombardment. We ascertained that the fluorescent proteins with specific targeting sequence to certain organelle was localized to proper organelle in various plant tissues and CFP and YFP were useful for two-color labeling of organelle. We showed that an Arabidopsis gene encoded the protein localized to both mitochondria and chloroplasts using this co-expression system. Targeting sequence of this gene predicted the localization of the protein to chloroplasts, but the part of targeting sequence from second exon predicted the localization to mitochondria. To examine whether these targeting sequences led the gene product to proper organelle, constructs joined to YFP behind each targeting sequence were prepared and expressed in Arabidopsis. The results showed the construct possessing whole targeting sequence of the gene was localized to chloroplasts, and the construct possessing the part of trageting sequence from second exon was localized to mitochondria.
植物化学調節学会の論文
2002-10-31

46 タンパク局在解析への改変緑色蛍光タンパクの利用

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