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园艺学报 ›› 2008, Vol. 35 ›› Issue (11): 1581-1586.

• 果树 • 上一篇    下一篇

龙眼胚胎F3H基因的cDNA克隆及序列分析

章希娟1,3,许鸿川1,2,游向荣1,2,李 燕1,2,陈清西1,3,陈 伟1,2*   

  1. 1福建农林大学蛋白质组学研究中心,福州 350002;2福建农林大学生命科学学院,福州 350002;3福建农林大学园艺学院,福州 350002)
  • 收稿日期:2008-08-12 修回日期:2008-10-13 出版日期:2008-11-25 发布日期:2008-11-25
  • 通讯作者: 陈 伟

Cloning and Sequence Analysis of F3H Gene cDNA from Longan Embryo

ZHANG Xi-juan1,3;XU Hong-chuan1,2;YOU Xiang-rong1,2;LI Yan1,2;CHEN Qing-xi1,3;CHEN Wei1,2*
  

  1. (1Research Center for Proteomics, Fujian Agriculture and Forestry University, Fuzhou 350002, China; 2College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China; 3College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou 350002, China)
  • Received:2008-08-12 Revised:2008-10-13 Online:2008-11-25 Published:2008-11-25
  • Contact: CHEN Wei

摘要:

采用IEF-SDS-PAGE双向电泳技术,对‘立冬本'龙眼合子胚发育不同时期的蛋白质进行分离,通过MALDI-TOF-TOF鉴定到其中一个上调差异表达的蛋白为黄烷酮-3-羟化酶 (flavanone 3-hydroxylase,F3H)。以‘立冬本’龙眼胚胎cDNA为模板,采用RT-PCR和RACE技术,获得F3H基因cDNA 1 404 bp全长序列。序列分析表明,F3H基因共编码365个氨基酸,其cDNA序列与柑橘、苹果、棉花等F3H基因的同源性均高达80%以上,该基因在GenBank中登录号为EF468104。

关键词: 龙眼, 胚胎, 黄烷酮-3-羟化酶, 基因克隆, 序列分析

Abstract:

The protein of 'Lidongben' longan zygotic embryos during different development stages were separated and compared by IEF-SDS-PAGE technique. Flavanone 3-hydroxylase was one of differential proteins identified by MALDI-TOF-TOF/MS. Then F3H gene cDNA sequence was cloned from longan zygotic embryos using RT-PCR and RACE technique. A 1 404 bp full-length cDNA sequence was obtained. Analysis of F3H gene cDNA indicated that it encoded a peptide containing 365 amino acids. The nucleotide sequence comparison with the F3H gene of Citrus sinensis, Gossypium hirsutum and Pyrus communi showed that identity was all higher than 80%. The sequence was accepted and released by GenBank (Accession number:EF468104)。

Key words: longan, embryo, flavanone 3-hydroxylase, gene cloning, sequence analysis

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