摘要
依据天麻(GastrodiaelataBl.f.flavidaS.Chow)抗真菌蛋白GAFP_1的N端部分氨基酸序列设计简并引物,通过RACE(快速分离cDNA末端)的方法扩增得到GAFP_1全长cDNA。该cDNA包含一个编码171个氨基酸的ORF,推导的多肽序列与测得的蛋白质部分序列相同;在5′端有一个长为55bp的5′非编码区;终止密码子下游有一个141bp长的3′非编码区,其中含有两个加poly(A)信号及长度为26个腺苷酸的poly(A)。经检索发现该推导蛋白序列与火烧兰(Epipactishelloborine)和二叶兰(Listeraovata)的甘露糖结合蛋白以及雪花莲(Galanthusnivalis)中的甘露糖结合凝集素具有很高同源性。
The cloning and sequencing of a full length cDNA of GAFP _1 ( Gastrodia antifungal protein), an antifungal protein from Gastrodia elata Bl. f. flavida S. Chow is reported. Degenerate primers were designed based on the N_terminal partial sequence from purified GAFP _1 to amplify the corresponding cDNA by rapid amplification of cDNA ends (RACE). A cDNA was obtained that contains an open reading frame for a peptide of 171 amino acids which matches the known peptide sequences. A 5′UTR (untranslated region) of 55 bp was found upstream from the translation initiation site. Two poly(A) adenylation sites were located downstream the stop codon. GAFP _1 cDNA and its deduced amino acid sequence share high homology with the mannose binding lectins from Epipactis helloborine , Listera ovata and snowdrop (Galanthus nivalis). The cDNA can now be used for testing the potential of GAFP _1 for engineering fungal resistance in crop plants.
基金
欧盟资助!(No .ERBIC1 8CT960 1 2 9)
关键词
天麻
抗真菌蛋白
快速分离
CDNA末端
Gastrodia eleta, Antifungal protein, Rapid amplification of cDNA ends (RACE), Cloning
