CAJ | 학술논문

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。 cD-NA全长1626 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ICE 1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。
이매화（ Prunus mume）품충‘삼륜옥접’위시재，근거평과（ Malus domestica） MdCIbHLH1기인서렬재매화기인조수거고중수색동원서렬병설계인물，통과PCR、RT-PCR획득ICE1기인적DNA、cDNA전장서렬。 cD-NA전장1626 bp，함유완정적개방열독광，편마541개안기산，기인편마구상존재3개내함자화4개외현자，함유bHLH구、부S구、핵정위신호구（ NLS）화전막구。 ExSAPy재선예측편마단백정친수성，아세포정위예측목적단백위우세포핵（98％）。 BLAST분석결과표명，기cDNA서렬급추도적안기산서렬균여평과、다（ Camellia sinensis）、첨양（Populus suaveolens）、지（Poncirus trifoliata）등물충적ICE1구유교고동원성，추단극륭도적cDNA가능시매화ICE 1기인，명명위PmICE1。실시형광정량분석결과표명，재4℃저온협박하，PmICE1기인표체량수협박시간적연장정상승추세，16 h시체도봉치，설명해기인재저온협박하정상조표체。
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) MdCIbHLH 1 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 PmICE1 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 suaveolens, and the cloned sequence may be a new member of ICE1 gene in Prunus mume.By relative real-time PCR experiment , the expression of PmICE1 gene increases at 4 ℃ at beginning and achieves the highest after 16 hours.Therefore, PmICE1 increases under low temperature stress.