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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 phenylalanine ammonia-lyase gene from Astragalus membranaceus. Methods PAL gene was cloned by RT-PCR and rapid amplification of cDNA ends using total RNA of Astragalus membranaceus as template. Results The cloned Astragalus phenylalanine ammonia-lyase gene was named AmPAL and Genbank accession number was EF567076. Sequence analysis showed that the full length of AmPAL was 2650bp and contained a complete open reading frame of 2154bp. AmPAL, a new member of the plant phenylalanine ammonia-lyase family, is presumed to encode a polypeptide of 718 amino acids, with a relative molecular mass of 7.805 × 104 and an isoelectric point of 5.96. AmPAL is closely related to the PAL amino acid sequence known to leguminous plants Homology and similarity are greater than 80%. Conclusion The PAL gene was successfully cloned from Astragalus membranaceus for the first time, which laid the foundation for the effective use of this gene in the regulation of phenylpropanoid metabolism in medicinal plants.