| 紫苏4-羟苯基丙酮酸双加氧酶基因
片段的克隆及表达分析 |
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| 引用本文： | 魏曼,吕晓玲,郝磊,宋西红,邬树伟.紫苏4-羟苯基丙酮酸双加氧酶基因
片段的克隆及表达分析[J].广东农业科学,2014,41(4):161-165. |
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| 作者姓名： | 魏曼 吕晓玲 郝磊 宋西红 邬树伟 |
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| 作者单位： | 天津科技大学食品工程与生物技术学院，天津300457 |
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| 基金项目： | 国家“863”计划项目（2007AA100401） |
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| 摘 要： | 为获得紫苏迷迭香酸合成途径旁路中的4-羟苯基丙酮酸双加氧酶（HPPD）基因，采用同源克隆的方法，
根据已报道的其他植物的HPPD 基因序列设计合成简并引物，克隆得到紫苏HPPD 基因片段，命名为PerHPPD-1，
该片段长663 bp，编码221 个氨基酸。通过氨基酸比对分析发现，其氨基酸序列与丹参和彩叶草的HPPD 基因片段
一致性分别为66.36%和77.93%，系统进化树分析又进一步表明该片段与唇形科植物的亲缘关系最近。荧光实时定
量PCR 检测结果显示，PerHPPD-1 基因在紫苏根、茎、叶中均有表达，在叶中表达量最高，其次为茎、根。激素（赤霉
素、茉莉酸甲酯、脱落酸）处理可在一定程度上上调PerHPPD-1 在叶中的表达水平。
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| 关 键 词： | 紫苏 迷迭香酸 4-羟苯基丙酮酸双加氧酶 序列分析 基因表达 |
Molecular cloning and expression analysis of 4-Hydroxyphenyl
pyruvate dioxygenase gene fragment from Perilla frutescens |
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| Abstract: | To obtain 4-Hydroxyphenyl pyruvate dioxygenase (HPPD) gene involved in the bypath of rosmarinic acid
(RA) biosynthesis pathway from Perilla frutescens, we designed degenerate primers according to parallel analysis of the
amino acid sequence of HPPD genes from other species and successfully cloned the fragment of HPPD gene by homology
cloning method. The fragment of HPPD designated as PerHPPD -1 was 663 bp, encoded 221 amino acids. Sequence
alignment revealed that the deduced amino acid sequence of PerHPPD -1 was 66.36, 77.93% identical to Salvia
miltiorrhiza, Coleus blume. Phylogenetic tree analysis showed that PerHPPD -1 had closest relationship with Lamiaceae
plants. The quantitative fluorescence Real-Time PCR analysis showed that the constitutive expression of PerHPPD-1 in
leaf was much higher than that in stem and root. Further expression analysis indicated that the hormone, including
Gibberellins, MeJA, ABA treatments could up-regulate the PerHPPD-1 level in leaves. |
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| Keywords: | Perilla frutescens rosmarinic acid 4-Hydroxyphenyl pyruvate dioxygenase sequence analysis gene
expression |
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