Abstract:

In this study, the full-length (1 870 bp) cDNA sequence of an ERF transcription factor genes, BgERF (GenBank accession No: GU593720) was isolated from Bruguiera gymnorhiza living in Shenzhen Mangrove Nature Reserves, using RACE-PCR technology. Bioinformatics analysis showed that BgERF had a complete open reading frame of 1 425 bp, encoding a protein of 475 amino acids. Three-dimensional structure was composed of a typical α-helix and 3 β-turn, with a conservative AP2/EREBP domain, and the 14th amino acid of AP2/EREBP domain is alanine (A). The 19th is aspartic acid (D), belongs to the ERF transcription factor family of proteins in AP2/EREBPF; the NCBI-BLAST homology sequence comparison and phylogenetic tree analysis showed that the BgERF had high homology in amino acid conserved regions with ERFs from most other plants like carrot, Arabidopsis, rice, pepper, tomato, soybean, potato., Gossypiumhirsutum L. This study also constructed a highly efficient plant expression vector PBI121-35s-BgERF, so as to lay a foundation for its functional and salt resistance mechanism research analysis.

Key words: Bruguiera gymnorhiza, RACE-PCR technology, AP2/EREBPF protein, ERF transcription factor, bioinformatics

摘要：

利用RACE-PCR技术,以深圳红树自然保护区的木榄为材料,克隆出一个ERF类转录因子基因的cDNA全长序列,命名为BgERF(GenBank序列号:GU593720)。生物信息学分析表明:该基因全长1 870 bp,完整开放阅读框1 425 bp,具有一个保守的AP2/EREBP结构域,且编码的由475个氨基酸组成的蛋白具有典型的1个α螺旋,3个β转角的三维结构,在AP2/EREBP结构域的第14位为丙氨酸(A),第19位为天冬氨酸(D),属于AP2/EREBPF蛋白家族中的ERF类转录因子；NCBI-BLAST比对结果表明:该基因与胡萝卜、拟南芥、水稻、辣椒、番茄、大豆、马铃薯、陆地棉等植物ERF类转录因子具有较高的同源性；并构建了高效植物表达载体PBI121-35s-BgERF,获得的转基因烟草与野生型相比,表现出明显的耐盐性,为该基因功能的深入分析和耐盐机理的研究奠定了基础。

关键词: 木榄；RACE-PCR技术；AP2/EREBPF蛋白；ERF转录因子；生物信息学