食品科学 ›› 2012, Vol. 33 ›› Issue (17): 145-149.

• 生物工程 • 上一篇    下一篇

滁菊类黄酮3′-羟化酶基因的克隆及分子特性

蔡 华,许雪峰,诸立新,孙艳辉,代洪亮,崔 勇   

  1. 滁州学院生物与食品工程学院
  • 收稿日期:2011-07-08 修回日期:2012-07-27 出版日期:2012-09-15 发布日期:2012-11-09
  • 通讯作者: 蔡华 E-mail:chczh@163.com
  • 基金资助:

    平行因子法和二维相关荧光光谱技术解析蛋白氧化体系;中国碧翠凤蝶的种群遗传结构及分子系统地理学研究;滁菊类黄酮3-羟化酶基因的克隆及其分子标记发掘

Cloning and Molecular Characterization of f3′h Gene in Dendranthema morifolium

  • Received:2011-07-08 Revised:2012-07-27 Online:2012-09-15 Published:2012-11-09

摘要: 以安徽滁州地区特有的药食两用植物资源——滁菊(Dendranthema morifolium‘chuju’)为试材,采用同源基因克隆方法克隆滁菊类黄酮3′-羟化酶基因(flavonoid 3′-hydroxylase,f3′h),NCBI注册号HQ256697。结果表明:采自不同地点的滁菊样品均能扩增出一条长381bp的f3′h基因片段,该片段与探针序列GU249553第3外显子对应区域的核苷酸序列同源性达98.69%,同源区域内有5个单核苷酸多态性位点(single nucleotide polymorphisms,SNP),其中3个SNPs导致氨基酸编码序列的改变。滁菊与葡萄属(Vitis vinifera)、高粱属(Sorghum bicolor)、芸苔属(Brassica napus)、牵牛属(Ipomoea nil Magenta)、苹果属(Malus×domestica)、番薯属(Ipomoea purpurea)等物种f3′h基因的核苷酸序列和氨基酸序列同源性分别在60%~98%和68%~97%之间。不同植物f3′h基因的分类与物种间的亲缘关系存在明显的相关性,菊属植物f3′h基因在系统进化中处于较高位置。

关键词: 滁菊, 类黄酮, 类黄酮3′-羟化酶基因, 外显子3, 序列分析

Abstract: Dendranthema morifolium ‘chuju’, a traditional Chinese medicine resource unique to Chuzhou, Anhui, was used to clone flavonoid 3′-hydroxylase (f3′h) gene with a registration number of HQ256697 in NCBI by homology-based cloning strategy in this study. The results showed that a separate 381 bp DNA fragment was amplified by polymerase chain reaction (PCR) from three‘chuju’ samples collected from different places. Sequence analysis showed that the amplified DNA fragments were closely homologous to the 3rd exon of f3′h gene in Chrysanthemum morifolium (GU249553) at a homology rate of 98.69% for nucleotide sequences. Five SNPs in the homology area were observed, and only 3 of them resulted in amino acid sequence variations. The homology rates of nucleotide and amino acid sequences between Dendranthema morifolium‘chuju’and Sorghum bicolor, Brassica napus, Ipomoea nil Magenta, Malus domestica or Ipomoea purpurea were 60%~98% and 68%~97%, respectively. The classification of f3′h gene in different plants was significantly related to their genetic relationship. The f3′h gene of Dendranthema was more advanced than that ofother plants in system evolution.

Key words: Dendranthema morifolium‘chuju’, flavonoid, f3′h gene, the 3rd exon, sequence analysis

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