Molecular cloning, in vitro expression and enzyme activity analysis of violaxanthin de-epoxidase from Oryza sauva L. |
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Authors: | Rongcheng Lin Liangbi Li Tingyun Kuang |
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Institution: | (1) Photosynthesis Research Center, Institute of Botany, Chinese Academy of Sciences, 100093 Beijing, China |
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Abstract: | The violaxanthin de-epoxidase gene was cloned from rice (Oryza sauva subsp japonica). The full length of the cDNA is 1887 bp, encoding a 446-amino acids protein with the transit peptide of 98 amino acids. The
bacterial expression vector pET-Rvde was constructed and the expression quantity of the exogenous protein increased with the
induction time by 0.4 mmol/L IPTG. Its molecular weight was similar with that of the native VDE. Western blotting indicated
that the expressed protein has immunological reaction with the VDE polyclonal antibody. The absorbance spectrum together with
xanthophyll pigments quantification by HPLC demonstrated that the expressed VDE has its enzyme activity, which can de-epoxidate
violaxanthin into antheraxanthin and zeaxanthin in vitro. |
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Keywords: | Oryza sativa L violaxanthin de-epoxidase clone expression enzyme activity |
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