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结合RACE和Genome Walking技术从十倍体长穗偃麦草中克隆了AP2家族的一个全长cDNA序列,命名为Ee-AP2.2。序列分析表明,该基因具有一个837bp的开放阅读框,编码279个氨基酸残基,含有一个保守的AP2结构域,是AP2大家族的一个新成员。该基因编码的氨基酸序列与GenBank已有的普通小麦AP2家族两个同源基因编码蛋白TaDREB1和TaDREBW50(登录号分别为:AAL01124.1和AAY44605.1)具有98%的氨基酸序列一致性,与大麦AP2蛋白HvDREB1-a(登录号AAY25517.1)、高羊茅AP2蛋白FaDREB2A(登录号CAG30547.1)及水稻OsDREB2.2(登录号AY064403)的氨基酸序列一致性分别为93%、86%和69%。说明该基因与小麦AP2家族基因的同源性最高。本研究除获得了长穗偃麦草一个重要抗逆转录因子基因EeAP2.2的全长cDNA序列外,也提供了一种快速、有效克隆功能基因的方法。
A full-length cDNA sequence of AP2 family was cloned from ten-day-old Agropyron elongatum with RACE and Genome Walking technology and named as Ee-AP2.2. Sequence analysis showed that the gene has an open reading frame of 837bp, encoding 279 amino acid residues and a conserved AP2 domain, which is a new member of AP2 family. The deduced amino acid sequence of this gene has 98% amino acid sequence identity with the two homologous genes AP2 family of TaDREB1 and TaDREBW50 (accession numbers: AAL01124.1 and AAY44605.1, respectively) existing in GenBank, The amino acid sequence identities of AP2 proteins HvDREB1-a (accession number AAY25517.1), tall fescue AP2 protein FaDREB2A (accession number CAG30547.1) and rice OsDREB2.2 (accession number AY064403) were 93%, 86% and 69 %. This gene has the highest homology with wheat AP2 family genes. In addition to obtaining the full-length cDNA sequence of EeAP2.2, an important anti-retroviral transcription factor gene of Erigeron elongatus, this study also provides a rapid and effective cloning of functional genes.