| 葡萄花青素还原酶基因ORF的克隆与原核表达 |
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| 引用本文： | 黄艺宁,柯丽娜,程祖锌.葡萄花青素还原酶基因ORF的克隆与原核表达[J].贵州大学学报(农业与生物科学版),2011,30(6):533-537. |
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| 作者姓名： | 黄艺宁 柯丽娜 程祖锌 |
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| 作者单位： | 漳州职业技术学院,福建漳州,363000 |
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| 摘 要： | 花青素还原酶是植物产生原花青素的一个重要基因。本研究以高原花青素葡萄品种"br"的叶片为材料,用同源克隆的方法克隆了原花青素合成关键酶花青素还原酶(anthocyanidin reductase,ANR)基因的开放阅读框ORF。结果得到1 017 bp的核苷酸序列,与葡萄(Vitis vinifera)、茶树(Camellia sinensis)ANR基因的同源性分别为99%、95%。将获得的开放阅读框与原核表达载体pET-28a构建成重组子pET-28a-ANR,并转化大肠杆菌BL21。SDS-PAGE电泳结果表明该重组子表达出预期大小(约45kDa)的融合蛋白。
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| 关 键 词： | 葡萄 原花青素 抗氧化 基因克隆 原核表达 |
The Cloning of Grape ANR Gene and its Prokaryotic Expression |
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| HUANG Yi-ning
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KE Li-na
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CHENG Zu-xin.The Cloning of Grape ANR Gene and its Prokaryotic Expression[J].Journal of Mountain Agriculture & Biology,2011,30(6):533-537. |
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| Authors: | HUANG Yi-ning KE Li-na CHENG Zu-xin |
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| Institution: | ( Zhangzhou Institue of Professional Technology, Zhangzhou Fujian 363000, China) |
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| Abstract: | Anthocyanidin Reductase gene （ANR） is well-known for its role in the synthesis of plant proanthocyanidin （PC）. In this study, grape cuhivar ＂br＂ was used to clone the open reading frame （ORF）_of anthoeyanidin reduetase gene （ANR） via homology method, and a 1 017 bp ORF was obtained. Alignment showed that the similarities between the target sequence and those of Vitis vinifera and Camellia sinensis were 99% and 84% , respectively. The recombinant pET-28a-ANR was constructed with the ORF into pET-28a. The plasmid was expressed in E. coil BL21 and the expected fusion protein with 45 kDa was detected using SDS-PAGE. |
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| Keywords: | grapes Proanthocyanins antioxidant gene cloning Prokaryotic expression |
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