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茶树紫黄素脱环氧化酶基因的体外定点突变及其突变体的表达和活性鉴定 被引量:6
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作者 韦朝领 江昌俊 +1 位作者 陶汉之 宛晓春 《中国生物化学与分子生物学报》 CAS CSCD 北大核心 2004年第1期73-78,共6页
经Swiss Prot蛋白质数据库中查询发现茶树紫黄素脱环氧化酶 (VDE)蛋白结构中含有lipocalin特征区 ,利用重叠延伸法把该特征区中最保守的Gly(GGG)和Trp(TGG)分别定点突变为Leu(TTG)和Tyr(TAG) .构建了表达载体pET 32a G→L和pET 32a W→... 经Swiss Prot蛋白质数据库中查询发现茶树紫黄素脱环氧化酶 (VDE)蛋白结构中含有lipocalin特征区 ,利用重叠延伸法把该特征区中最保守的Gly(GGG)和Trp(TGG)分别定点突变为Leu(TTG)和Tyr(TAG) .构建了表达载体pET 32a G→L和pET 32a W→Y ,在E .coliBL2 1trxB(DE3)进行了诱导表达 ,并对表达蛋白进行His Tag亲和纯化 .结果表明 ,表达蛋白的分子量和预计的相等 ,为 5 8 9kD ,表达量占菌体总蛋白的 4 5 % .体外酶促反应分析得出 ,G→L或W→Y突变都能导致茶树VDE生物活性大幅度降低 ,证明了lipocalin特征区是VDE的主要活性中心之一 . 展开更多
关键词 茶树 紫黄素脱环氧化酶 定点突变 表达 生物活性
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植物紫黄素脱环氧化酶结构特征的生物信息学分析 被引量:4
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作者 韦朝领 江昌俊 +1 位作者 陶汉之 宛晓春 《安徽农业大学学报》 CAS CSCD 北大核心 2002年第3期250-255,共6页
利用国际互联网上的生物学数据库 (Gen Bank,Swiss-Prot和 Prosite)所提供的信息资源和工具(PHDsec和 Signal P)及软件技术 (DNAstar) ,从生物信息学角度对植物紫黄素脱环氧化酶 (VDE)进行序列的比较分析和结构预测 ,得出了其转运肽特... 利用国际互联网上的生物学数据库 (Gen Bank,Swiss-Prot和 Prosite)所提供的信息资源和工具(PHDsec和 Signal P)及软件技术 (DNAstar) ,从生物信息学角度对植物紫黄素脱环氧化酶 (VDE)进行序列的比较分析和结构预测 ,得出了其转运肽特征、相关保守位点和特殊结构域以及序列的二级结构特征 。 展开更多
关键词 紫黄素脱环氧化酶 生物信息学 序列分析 结构域 黄素 植物光保护机制
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Xanthophyll Cycle and Its Molecular Mechanism in Photoprotection 被引量:10
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作者 林荣呈 许长成 +1 位作者 李良璧 匡廷云 《Acta Botanica Sinica》 CSCD 2002年第4期379-383,共5页
When plants absorb more light than that can be used for photosynthesis, the excessive energy can cause photoinhibition and even photooxidation of photosynthetic apparatus. Xanthophyll cycle-dependent photo-protection ... When plants absorb more light than that can be used for photosynthesis, the excessive energy can cause photoinhibition and even photooxidation of photosynthetic apparatus. Xanthophyll cycle-dependent photo-protection is believed to be the main mechanism for plants to deal with excessive light energy. This review focuses on molecular biological aspects and regulations of violaxanthin de-epoxidase and zeaxanthin epoxidase involved in xanthophyll cycle. We will summarize the functions of xanthophyll cycle, especially recent advances in its thermal dissipation mechanism of photoprotection. Some interesting issues deserving further study will be discussed. 展开更多
关键词 xanthophyll cycle violaxanthin de-epoxidase thermal dissipation PHOTOPROTECTION
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Changes in Violaxanthin Deepoxidase Activity and Unsaturation of Thylakoid Membrane Lipids in Indica and Japonica Rice Under Chilling Condition and Strong Light 被引量:1
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作者 季本华 曹云英 +3 位作者 谢焕松 朱素琴 马强 焦德茂 《Acta Botanica Sinica》 CSCD 2003年第9期1063-1070,共8页
To explore the differences of sensitivities to chilling and strong light in indica and japonica rice (Oryza sativa L), the changes in unsaturation of thylakoid membrane lipids and xanthophyll cycle were studied under ... To explore the differences of sensitivities to chilling and strong light in indica and japonica rice (Oryza sativa L), the changes in unsaturation of thylakoid membrane lipids and xanthophyll cycle were studied under chilling condition and strong light. The contents of unsaturated fatty acids of thylakoid membrane lipids decreased and that of the saturated ones increased with the time of chilling and strong light treatment, resulting in the reduction of the index of unsaturation of fatty acids (IUFA). The activity of violaxanthin deepoxidase (VDE), a key enzyme of xanthophyll cycle, also reduced. The content of violaxanthin (V) increased, and the contents of antheraxanthin (A) and zeaxanthin M decreased, the ratio of (A+Z)/ (A+Z+V) decreased correspondingly. Arrhenius analysis showed that VDE was sensitive to both chilling and unsaturation level of thylakoid membrane lipids. Correlation analysis showed that there was distinctly positive relationships between IUFA of thylakoid membrane lipids and the activity of VDE, Fv/Fm, and D, protein content. Lower IUFA values, less fluidity and stability of thylakoid membrane lipids, lower VDE activity and (A+Z)/(A+Z+V) ratio were found in indica rice cv. Shanyou 63 than in japonica rice cv. 9516 under chilling and strong light. 展开更多
关键词 thylakoid membrane lipids xanthophyll cycle violaxanthin deepoxidase Arrhenius plotting RICE
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Cloning and Expression Analysis of Violaxanthin De-Epoxidase (VDE) cDNA in Wheat 被引量:1
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作者 张吉军 英加 +2 位作者 常胜合 李滨 李振声 《Acta Botanica Sinica》 CSCD 2003年第8期981-985,共5页
Violaxanthin de-epoxidase (VDE) is the key enzyme in the xanthophyll cycle and protects plant photosynthetic apparatus from the damage of excessive light. A wheat (Triticum aestivum L cv. Xiaoyan 54) VDE cDNA was obta... Violaxanthin de-epoxidase (VDE) is the key enzyme in the xanthophyll cycle and protects plant photosynthetic apparatus from the damage of excessive light. A wheat (Triticum aestivum L cv. Xiaoyan 54) VDE cDNA was obtained using RT-PCR method. Its deduced protein sequence shares high identity with that of Arabidopsis and rice. Southern blot revealed that there are three copies of VDE gene per haploid genome of wheat. VDE transcript levels were higher in green leaf than in root, seed and etiolated leaf. Northern blotting analysis indicated that VDE mRNA level is induced during greening process of etiolated wheat seedling and increased by intense light illumination. 展开更多
关键词 violaxanthin de-epoxidase xanthophylls cycle PHOTOPROTECTION Triticum aestivum
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