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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (24): 207-212.

所属专题： 生物技术

• 生物技术科学 • 上一篇    下一篇

紫苏酪氨酸氨基转移酶基因片段的克隆及表达分析

吕晓玲 郝磊 王芳 黄晨   

  • 收稿日期:2011-12-09 修回日期:2012-02-20 出版日期:2012-08-25 发布日期:2012-08-25
  • 基金资助:

    国家高技术研究发展计划（863计划）

Molecular Cloning and Expression Analysis of Tyrosine Aminotransferase Gene Fragment in Perilla frutescens

  • Received:2011-12-09 Revised:2012-02-20 Online:2012-08-25 Published:2012-08-25

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摘要：

为获得紫苏迷迭香酸合成途径中的酪氨酸氨基转移酶基因，本研究采用同源克隆的方法，根据已报道的其他物种的TAT基因序列设计并合成简并引物，成功克隆得到了紫苏TAT基因片段（GenBank登录号：JN032113.1），该片段长为579 bp，共编码193个氨基酸残基，并命名为PfTAT。氨基酸序列比对分析发现其与彩叶草、丹参、拟南芥和罂粟的一致性分别为97%、94%、69%和58%，系统进化树分析表明PfTAT与唇形科植物的亲缘关系最近。采用半定量RT-PCR法分析PfTAT在紫苏的根、茎、叶中均有表达，且叶中的表达量较高，内源性植物激素信号分子对PfTAT表达量影响的实验表明，脱落酸、水杨酸处理均能够不同程度得上调PfTAT转录水平的表达。

关键词: 致病基因, 致病基因

Abstract:

The purposes of this paper was to obtain tyrosine aminotransferase (TAT) gene involved in rosmarinic acid (RA) biosynthesis pathway from Perilla frutescens. According to a parallel analysis of the amino acid sequence of TAT genes from other species, degenerate primers were designed and the fragment of TAT gene in Perilla frutescens was successfully cloned by homology cloning method (GenBank accession No. JN032113.1), and gene fragment was 579 bp encoding 193 amino acid protein. Sequence alignment revealed that the deduced amino acid sequence of PfTAT was 97%, 94%, 69% and 58% identical to Coleus blumei, Salvia miltiorrhiza, Arabidopsis thaliana and Papaver somniferum, respectively. Phylogenetic tree analysis showed that PfTAT had the closest relationship with Lamiaceae plants. The PfTAT expressed in all the tested tissues but at different levels with higher expression in leaf using the semi-quantitative RT-PCR analysis. Further expression analysis reveals that the signaling components such as abscisic acid (ABA) and salicylic acid (SA) up-regulate the PfTAT transcript levels in different degree.

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