桃果实乙烯反应因子PpERF1基因的克隆及序列分析

Cloning and Sequencing Analysis of PpERF1 Gene from Peach Fruit

以成熟肥城桃果实的cDNA为模板,根据EST库中的核苷酸序列,设计2条特异引物分别与B26进行PCR扩增,结果得到了该基因下游包括3′端非编码区的765bp大小的cDNA片段。PpERF1基因全长为1263bp,包含一个708bp大小的完整开放阅读框(ORF)。序列分析表明,PpERF1与番茄、拟南芥、苹果和葡萄等在DNA结合功能域的约57个氨基酸中,同源性可达68.4%～100%,且在该区域中存在1个α-螺旋和3个反平行链构成的β-折叠结构。经同源性分析,该序列与拟南芥、葡萄、豌豆等植物的氨基酸同源性为58.8%～67.5%,属于同一类ERF家族成员。

According to the nucleic acid sequence from EST database, two specific primers were designed and a 765 bp eDNA fragment containing 3＇ untranslated region was amplified by RT-PCR from ripening peach fruit. The cDNA, named PpERF1, contains an ORF （open reading frame） which is 708 bp in length. PpERF1 contains a highly conserved, plant-specific DNA-binding domain, which is composed of one α-helix and. three 13-sheets Multiple alignment of the ERF conserved domains shows that PpERF1 has 68.4%-100% identity to ERF homolog in other plants. PpERF1 shared high similarity at both nucleotide and polypeptide level with corresponding sequences from Arabidopsis, grape and pea.