紫苏酪氨酸氨基转移酶基因片段的克隆及表达分析  

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

作　　者：吕晓玲[1] 郝磊[1] 王芳[1] 黄晨[2] 

机构地区：[1]天津科技大学,天津300457 [2]天津出入境检验检疫局,天津300461

出　　处：《中国农学通报》2012年第24期207-212,共6页Chinese Agricultural Science Bulletin

基　　金：国家高技术研究发展计划("863"计划)"生物酶法食品添加剂和配料高效安全制造"(2007AA100401)

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579bp,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。The purposes of this paper was to obtain tyrosine aminotransferase （TAt） gene involved in rosmarinic acid （RA） biosynthesis pathway from Perilla fruteseens. 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 fruteseens 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 PtTATwas 97%, 94%, 69% and 58% identical to Coleus blumei, Salvia miltiorrhiza, Arabidopsis thaliana and Papaver somnifemm, respectively. Phylogenetic tree analysis showed that PiTAT had the closest relationship with Lamiaeeae plants. The PtTA T 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 revealsed that the signaling components such as abscisic acid （ABA） and salicylic acid （SA） up-regulated the PftTAT transcript levels in different degree.

关 键 词：紫苏 迷迭香酸 酪氨酸氨基转移酶 基因克隆 序列分析 表达 

分 类 号：Q78[生物学—分子生物学]