Cloning and Expression of a Seed-Specific Metallothionein-Like Protein from Sesame
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概要
- 論文の詳細を見る
A cDNA clone, SiMT encoding an Ec type of metallothionein (MT)-like protein, was isolated from maturing seeds of sesame (Sesamum indicum L.), and its deduced protein sequence shared 47–65% similarity to other known Ec type of MT-like proteins with three highly conserved cysteine-rich segments. The transcript of SiMT was exclusively accumulated in maturing seeds from two weeks after flowering to the end of seed maturation. The results of a southern blot analysis suggested that one SiMT and one SiMT-like gene were present in the sesame genome. Recombinant SiMT fused with glutathione-S-transferase (GST) was over-expressed in Escherichia coli, and purified to homogeneity by affinity chromatography. Recombinant SiMT released from GST was harvested after factor Xa cleavage. Migration of the recombinant SiMT during SDS–PAGE was accelerated when its binding metal ions were depleted by EDTA. The metal-binding capability of recombinant SiMT was measured by inductively-coupled plasma atomic emission spectrometry. Our results show that the recombinant SiMT could trap zinc or copper ions, but not manganese ions, with a stoichiometric ratio (metal ion/SiMT) of approximately 2.
- 社団法人 日本農芸化学会の論文
- 2005-12-23
著者
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TZEN Jason
Graduate Institute of Agricultural Biotechnology National Chung-Hsing university
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Tzen J
National Chung Hsing Univ. Taichung Twn
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Tzen Jason
Graduate Inst. Of Biotechnology National Chung Hsing Univ.
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CHYAN Chia-Lin
Department of Chemistry, National Dong Hwa University
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LEE Tiger
Graduate Institute of Biotechnology, National Chung Hsing University
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LIU Chiung-Pin
Division of Watershed Management, Taiwan Forestry Research Institute
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YANG Yuan-Chang
Department of Biotechnology, National Formosa University
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CHOU Wing-Ming
Department of Biotechnology, National Formosa University
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Chyan Chia-lin
Department Of Chemistry National Dong Hwa University
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Lee Tiger
Graduate Institute Of Biotechnology National Chung Hsing University
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Liu Chiung-pin
Division Of Watershed Management Taiwan Forestry Research Institute
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Chou W‐m
Department Of Biotechnology National Formosa University
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Yang Yuan-chang
Department Of Biotechnology National Formosa University
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Chou Wing-ming
Department Of Biotechnology National Formosa University
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