摘要
以梅花(Prunus mume)品种‘三轮玉蝶’为试材,根据苹果(Malus domestica)MdCIbHLH1基因序列在梅花基因组数据库中搜索同源序列并设计引物,通过PCR、RT-PCR获得ICE1基因的DNA、cDNA全长序列。cDNA全长1 626 bp,含有完整的开放阅读框,编码541个氨基酸,基因编码区上存在3个内含子和4个外显子,含有bHLH区、富S区、核定位信号区(NLS)和转膜区。ExSAPy在线预测编码蛋白呈亲水性,亚细胞定位预测目的蛋白位于细胞核(98%)。BLAST分析结果表明,其cDNA序列及推导的氨基酸序列均与苹果、茶(Camellia sinensis)、甜杨(Populus suaveolens)、枳(Poncirus trifoliata)等物种的ICE1具有较高同源性,推断克隆到的cDNA可能是梅花ICE1基因,命名为PmICE1。实时荧光定量分析结果表明,在4℃低温胁迫下,PmICE1基因表达量随胁迫时间的延长呈上升趋势,16 h时达到峰值,说明该基因在低温胁迫下呈上调表达。
A putative sequence of ICE1 gene was identified in the genome database of Prunu mume according to the ICE1 tran- scription factor of Apple （Malus domestica） MdCIbHLH1 gene. ICE1 gene was isolated from Prunus mume ＇San Lun Yu Die＇ by PCR and RT-PCR. A 1 626-bp cDNA clone was obtained including a putatively entire open reading frame （ORF） and encodes a protein of 541 amino acids. The coding regions of the gene include three introns and four exons. The predic- ted PmICEI protein contains a typical bHLH domain, a serine-rich region （S-rich） , a nuclear localization signal （NLS）, a transmembrane domain, N-glycosylation and kinase phosphorylation sites. ExSAPy online predictive coding protein is hy- drophilic. The possibility of location in the nucleus is 98% according to the subcellular localization prediction. By bioin- formatics analysis, the cDNA is highly homologous to ICE1 from MaluS domestica, Camellia sinensis, Poncirus trifoliata and Populus suaveolen,~, and the cloned sequence may be a new member of ICE1 gene in Prunus mume. By relative real- time PCR experiment, the expression of PmlCE1 gene increases at 4 ℃ at beginning and achieves the highest after 16 hours. Therefore, PmICEI increases under low temperature stress.
出处
《东北林业大学学报》
CAS
CSCD
北大核心
2014年第4期21-25,共5页
Journal of Northeast Forestry University
基金
国家“863”计划项目(2011AA100207、2013AA102607)
