| 草莓八氢番茄红素合成酶基因的克隆及其表达特性 |
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| 引用本文： | 朱海生,李永平,温庆放.草莓八氢番茄红素合成酶基因的克隆及其表达特性[J].中国农业科学,2011,44(2):349-357. |
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| 作者姓名： | 朱海生 李永平 温庆放 |
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| 作者单位： | (福建省农业科学院作物研究所/福建省农业科学院蔬菜研究中心/福建省蔬菜工程技术研究中心); |
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| 基金项目： | 福建省科技重大专项(2008NZ0002); 国家科技支撑项目(2007BAD07B03); 福建省自然科学基金项目(2010J0111); 福建省属公益类科研院所基本科研专项(2010R1028-5); 福建省农业科学院博士科研启动基金项目(BS04) |
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| 摘 要： | 【目的】分离和克隆草莓果实八氢番茄红素合成酶基因psy,分析其序列特征,了解其在不同组织部位的表达情况。【方法】采用RT-PCR和RACE技术从草莓果实中克隆草莓类胡萝卜素合成途径中关键基因psy,用生物信息学方法对获得的cDNA序列及推定氨基酸序列进行分析,并用半定量PCR法研究psy基因在不同组织中的表达。【结果】分离到psy基因,GenBank登录号为FJ784889。该cDNA全长1 458 bp,具有1个1 194 bp的完整开放阅读框(ORF),编码398个氨基酸。序列分析表明,psy编码的氨基酸序列与其它植物的PSY蛋白有很高的相似性。系统进化树分析显示,草莓PSY与胡萝卜和玉米的PSY蛋白亲缘关系比较近。原核表达结果表明psy基因在大肠杆菌中获得高效表达。利用半定量RT-PCR技术进行组织表达模式分析发现,psy基因在草莓的花、叶片和果实中均有表达。表达量为花＞红果＞粉红果＞白果＞青果＞老叶＞新叶。【结论】从草莓中克隆到类胡萝卜素生物合成的关键酶基因psy,该基因可能参与调控类胡萝卜素的合成。
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| 关 键 词： | 草莓 八氢番茄红素合成酶 psy 基因克隆 |
| 收稿时间： | 2010-06-29; |
Cloning and Expression Analysis of Phytoene Synthase Gene in Fragaria ananassa Duch. |
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| ZHU Hai-sheng,LI Yong-ping,WEN Qing-fang.Cloning and Expression Analysis of Phytoene Synthase Gene in Fragaria ananassa Duch.[J].Scientia Agricultura Sinica,2011,44(2):349-357. |
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| Authors: | ZHU Hai-sheng LI Yong-ping WEN Qing-fang |
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| Affiliation: | ZHU Hai-sheng,LI Yong-ping,WEN Qing-fang (Crops Research Institute,Fujian Academy of Agricultural Sciences/Vegetable Research Center,Fujian Academy of Agricultural Sciences/Fujian Engineering Research Center for Vegetables,Fuzhou 350013) |
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| Abstract: | 【Objective】The aim of this study was to clone full-length cDNA of key enzyme gene psy for carotene biosynthesis in strawberry, to investigate its sequence characteristics and to analyze its expression in different organ.【Result】 The psy gene cDNA sequence was cloned from strawberry fruit using RT-PCR and RACE techniques, bioinformatics methods were used to analyze cDNA sequence obtained and putative amino acid sequence,Semi-quantitative PCR method were used to study the expression of psy gene in different tissues. 【Result】 The full-length cDNA of psy (GenBank accession number：FJ784889) in strawberry was cloned, The sequence consists of 1 458 bp with an intact open reading frame of l 194 bp, encoding a polypeptide of 398 amino acids. Homology analysis showed that the deduced PSY protein was highly homologous to other PSY proteins from different species. Phylogenetic analysis also indicated that PSY was more related to PSY of carrot and corn. Prokaryotic expression showed recombinant plasmid was efficiently expressed in Escherichia coli BL21. The semi-quantitative RT-PCR analysis revealed psy can be expressed in different strawberry tissues, and the expression level was differed in these tissues, the highest expression level was detected in flower, while moderate expression levels were found in fruits at the red ripening stage, fruits at the pink ripening stage, white fruits, green fruits and old leaf, the lowest level was present in new leaves. 【Conclusion】 Gene psy was firstly isolated and characterized from strawberry, which may participate in carotene biosynthesis. |
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| Keywords: | strawberry phytoene synthase psy gene cloning |
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