马铃薯乙烯响应因子基因的克隆表达及生物信息学分析

Cloning,bioinformatics and expression analysis of ethylene response factor gene of potato

根据番茄ERF5基因(NM_001247583.1)序列设计引物,采用RT-PCR方法得到马铃薯ERF基因的cDNA,并对其进行生物信息学及表达分析.结果表明:该cDNA全长789bp,阅读框为732bp,编码243个氨基酸.序列同源性分析表明该基因序列与番茄ERF5基因的序列同源性达82%,氨基酸序列同源性达70%.对该马铃薯ERF基因的氨基酸序列经BLAST进行多序列比对,发现该氨基酸序列的98～160bp为AP2结构域,属于AP2/EREBP转录因子的EREBP亚族ERF类.蛋白质三级结构预测表明该基因的AP2结构域非常保守.对ERF基因进行荧光定量PCR分析,表明该基因可能参与了马铃薯块茎解除休眠与发芽过程的调控.

Ethylene response factor（ERF） is a kind of transcription factors（TF） particularly represented in plant,which are involved in plant growth and development,hormone response,disease and insect resistance,and abiotic stress resistance.Primers were designed based on tomato ERF5 gene sequence（NM_001247583.1）.The cDNA of potato ERF gene with a total of 789 bp and open reading frame of 732 bp encoding a protein with 243 amino acids was obtained by RT-PCR method.Sequence alignment result showed that there was a similarity of 82% of ERF gene and 70% of amino acid between potato and tomato.The result from BLAST alignment of amino acid sequence of ERF gene demonstrated that a segment of 98~160 bp of amino acid was AP2 domain,and the potato ERF gene belonged to ERF group of EREBP subfamilies of AP2/EREBP transcription factors.Protein tertiary structure prediction indicated that the AP2 domain was much conserved.Fluorescence quantitative PCR analysis of ERF gene showed that it was probably taken part in potato tuber dormancy release and sprouting.