目的对膜荚黄芪苯丙氨酸解氨酶基因进行克隆及序列分析。方法应用RT-PCR和cDNA末端快速扩增法,以膜荚黄芪根总RNA为模板克隆PAL基因。结果所克隆的膜荚黄芪苯丙氨酸解氨酶基因命名为AmPAL,Genbank登录号为EF567076。序列分析表明,AmPAL全长为2650bp,含有一个完整的2154bp开放读码框。AmPAL是植物苯丙氨酸解氨酶家族的一个新成员,推测其编码718个氨基酸的多肽,相对分子质量为7.805×104,等电点为5.96,与豆科植物已知的PAL氨基酸序列的同源并且相似性都大于80%。结论首次成功地从膜荚黄芪中克隆出了PAL基因,为有效利用该基因调控药用植物苯丙烷代谢途径奠定了基础。

Objective To clone and sequence the cDNA encoding phenylalanine ammonia-lyase(PAL) gene from Astragalus membranaceus.Methods RT-PCR and RACE Techniques were used to clone a phenylalanine ammonia-lyase gene from A. membranaceus roots with the total RNA as the template. Resalts The cloned gene named as A mPAL and the Genbank registry number is EF567076.Squence analysis showed that the full-length of AmPAL cDNA was 2650 by，including a 2154 by open reading frame (ORF).A mPAL was a new number of PAL family that consisted of 718 amino acids with predicted molecular weight of 7.805×104 and isoelectric point(PI) of 5.96.At the same time，A mPAL had the homology with PAL of known leguminous plants and shared above 80%identity of amino acid sequences.Conclusion It is the first report that a novel PAL gene is clonedfrom A. embranaceus.This work lays afounlotion for regulating phenylpropanoid pathway of medical plant with A mPAL.

国家自然科学基金资助项目(30860036)