摘要
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 gene from Astragalus rnernbranaceus. Methods RT-PCR and RACE Techniques were use lyase （PAL） d to clone a phenylalanine ammonia-lyase gene from A. rnembranaceus roots with the total RNA as the template. Results The cloned gene named as ArnPAL and the Genbank registry number is EF567076. Squence analysis showed that the full-length of ArnPAL cDNA was 2 650 bp, including a 2 154 bp open reading frame （ORF）. ArnPAL was a new number of PAL family that consisted of 718 amino acids with prediated molecular weight of 7. 805×10^4 and isoelectric point （PI） of 5.96. At the same time, AmPAL had the homology with PAL of known leguminous plants and shared above 80% identity of amino acid sequences. Cola- clusion It is the first report that a novel PAL gene is cloned from A. membranaceus. This work lays a foundation for regulating phenylpropanoid pathway of medical plant with ArnPAL.
出处
《中草药》
CAS
CSCD
北大核心
2010年第3期456-460,共5页
Chinese Traditional and Herbal Drugs
基金
国家自然科学基金资助项目(30860036)
关键词
膜荚黄芪
苯丙氨酸解氨酶
基因克隆
Astragalus mernbranaceus （Fisch.） Bunge
phenylalanine ammonia-lyase
gene cloning
