| 芒果无机焦磷酸酶基因的克隆及其表达载体构建 |
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| 引用本文： | 白蓓蓓,荆永琳,蔡秉宇,蓝丽,王佳,赵志常.芒果无机焦磷酸酶基因的克隆及其表达载体构建[J].热带作物学报,2019,40(2):308-313. |
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| 作者姓名： | 白蓓蓓 荆永琳 蔡秉宇 蓝丽 王佳 赵志常 |
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| 作者单位： | 1. 海南大学热带农林学院, 海南海口 5702282. 中国热带农业科学院热带作物品种资源研究所/农业农村部华南作物基因资源与种质创制重点实验室, 海南儋州 571737 |
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| 基金项目： | 国家自然科学基金资助项目(31471850);中国热带农业科学院基本科研业务费项(1630032017005);中国热带农业科学院基本科研业务费项(1630032017004) |
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| 摘 要： | 无机焦磷酸酶（inorganic pyrophosphatase,PPase）催化焦磷酸（PPi）水解为2个无机正磷酸（Pi）,是蔗糖合成途径中的调控关键节点之一。本研究根据已经报道的PPase基因的序列设计兼并引物,采用3′ RACE和5′ RACE方法,从贵妃芒果的果实中克隆得到了一个芒果PPase基因,将其命名为MiPPase,其全长cDNA序列为1014 bp,开放阅读框为837 bp,编码278个氨基酸,分子量为30.85 ku,等电点为4.68。通过系统发育分析发现该基因编码的蛋白与红花烟草具有较近的亲缘关系。为深入探究MiPPase基因在蔗糖代谢过程中的作用,本研究成功构建出pGreenII 62-SK-MiPPase基因过量表达载体,为后续研究MiPPase基因在芒果果实蔗糖合成的作用机理提供理论依据。
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| 关 键 词： | 芒果 PPase 基因克隆 表达载体构建 |
| 收稿时间： | 2018-07-21 |
Cloning and Overexpression Vector Construction of Inorganic Pyrophosphatase Gene of Mangifera indica L. |
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| BAI Beibei,JING Yonglin,CAI Bingyu,LAN Li,WANG Jia,ZHAO Zhichang.Cloning and Overexpression Vector Construction of Inorganic Pyrophosphatase Gene of Mangifera indica L.[J].Chinese Journal of Tropical Crops,2019,40(2):308-313. |
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| Authors: | BAI Beibei JING Yonglin CAI Bingyu LAN Li WANG Jia ZHAO Zhichang |
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| Institution: | 1. Institute of Tropical Agriculture and Forestry, Hainan University, Haikou, Hainan 570228, China2. Tropical Crops Genetic Resources Institute, Chinese Academy of Tropical Agricultural Sciences / Key Laboratory of Crop Gene Resources and Germplasm Enhancement in Southern China, Ministry of Agriculture and Rural Affairs, Danzhou, Hainan 571737, China |
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| Abstract: | The hydrolysis of pyrophosphoric acid (PPi) is catalyzed by inorganic pyrophosphate phosphatase (PPase) into two inorganic orthophosphoric acid (Pi),which is one of the key regulatory nodes in the sucrose synthesis pathway. According to the reported sequence of PPase gene, primers were designed in this study. PPase gene was cloned in the fruit of Guifei mango using the methods of 3′ Race and 5′ Race, and named MiPPase. Its full-length of the cDNA sequence was 1014 bp and the open reading frame was 837 bp. It encoded 278 amino acids with molecular weight of 30.85 ku and its isoelectric point was 4.68. Phylogenetic analysis showed that the protein encoded by the PPase gene was closely related to Nicotiana tabacum. In order to investigate the role of MiPPase gene in sucrose metabolism, the pGreenII 62-SK-MiPPase gene overexpression vector was successfully constructed, which would provide a theoretical basis for the further study of the mechanism of MiPPase gene on the sucrose synthesis in mango fruit. |
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| Keywords: | Mangifera indica PPase gene cloning expression vector construction |
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