Variation of the Amino Acid Content of Arabidopsis Seeds by Expressing Soybean Aspartate Aminotransferase Gene
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概要
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We have increased the contents of several amino acids in the seeds of Arabidopsis thaliana by introduction of aspartate aminotransferase (AAT), an enzyme of the aspartate biosynthetic path-way. mRNA was prepared from one-week-old seedlings of Glycine max cv. enrei and the cDNA encoding AAT5 was isolated and linked to the CaMV35S promoter in the plant vector pBI121. The AAT5 gene encodes a protein of 462 amino acid residues that shows 51% amino acid sequence similarity to A. thaliana chloroplast Asp3. The soybean AAT5 also contains a chloroplast transit peptide and is able to functionally complement a Saccaromyces cerevisiae mutant lacking the Asp5 gene. A. thaliana was transformed with the AAT5 gene from Agrobacterium tumefaciens by the vacuum infiltration method. The AAT5 gene was detected in the transcript and genomic DNA from the transgenic T2 plants. The T3 progeny showed a 3:1 segregation ratio indicating the presence of a single integration. Expression of G. max AAT5 in A. thaliana transformants caused 3-, 4-, 23-, and 50-fold increases in the contents of free glycine, alanine, asparagine, and glutamine, respectively, in the T3 seeds. A decrease in the contents of valine, tyrosine, isoleucine, leucine, and phenylalanine by several folds was also observed. Thus, it is of interest that a key gene expression resulted in marked changes of metabolites in plant seeds.
- 社団法人日本生物工学会の論文
- 2002-09-25
著者
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HAYASHI Makoto
Department of Basic Medical Research and Education, Ehime University Graduate School of Medicine
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MUROOKA YOSHIKATSU
Department of Agricultural Chemistry, Faculty of Bioresources, Mie University, Department of Biotech
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Mori Y
Department Of Biotechnology Graduate School Of Engineering Osaka University
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Mori Yukiko
Department Of Pediatrics Fukui Red Cross Hospital
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Mori Yukiko
Department Of Biotechnology Graduate School Of Engineering Osaka University
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Murooka Yoshikatsu
Depart. Of Bioteclnology Graduate School Of Eng. Osaka Univ.
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Hayashi Makoto
Depart. Of Bioteclnology Graduate School Of Eng. Osaka Univ.
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HAYASHI MAKOTO
Department of Biotechnology, Graduate School of Engineering, Osaka University
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