摘要： 苯丙氨酸解氨酶(PAL) 是迷迭香酸合成途径中的关键酶之一,根据其他植物PAL基因的保守区域设计特异引物,利用3'-RACE-PCR技术,本研究首次从夏枯草中克隆得到了PAL基因的cDNA片段序列,命名为PvPAL,Gen-Bank登录号为JN65446。PvPAL基因cDNA片段长1 306 bp,其中编码区域为1 047 bp,编码349个氨基酸。蛋白质序列多重比较结果显示,PvPAL蛋白质序列与丹参、地黄、黄芩、藿香等植物的PAL蛋白质高度同源。PAL系统进化树分析结果表明,PvPAL与唇形科植物的PAL基因亲缘关系最近。组织表达分析结果显示,PvPAL基因在根、茎、叶中均表达,其中根中表达量最高。PvPAL基因的克隆为进一步研究夏枯草迷迭香酸合成的分子机制奠定了基础。

关键词: 夏枯草, 苯丙氨酸解氨酶, 基因克隆, 序列分析, 表达

Abstract: Phenylalanne ammonial-lyase(PAL) is one of the key enzymes involved in rosmarinic acid biosynthetic pathway. In this study, a PAL gene, named PvPAL, was cloned from Prunella vulgaris at the first time by 3'-RACE-PCR and using the specific primer, which was designed according to the homologus sequences of PAL genes from other plants. The GenBank accession number of PvPAL is JN65446. The length of PvPAL cDNA fragment is 1 306 bp, including a 1 047 bp-length coding sequence, which encoded a 349-amino-acid protein. Sequence multiple-alignment revealed that PvPAL protein had extensive homology with those of other plants as Salvia miltiorrhiza, Rehmannia glutinosa, Scutellaria baicalensis and Agastache rugosa. Phylogenetic tree analysis showed that PvPAL had closest relationship with PALs from Lamiaceae plants than from other plants. Tissue expression analysis indicated that PvPAL expressed in all tissues examined, but highest in roots. The isolation of PvPAL provided basis for further studying the molecular mechanism of rosmarinic acid biosynthesis in P. vulgaris.

Key words: Prunella vulgaris, Phenylalanne ammonial-lyase, Gene cloning: Sequence analysis, Expression

中图分类号: 

• Q78