CAJ | 학술논문

从鹰嘴豆中克隆生物节律钟 LHY 基因的 cDNA 全长序列,进行序列信息学分析.通过同源克隆策略,利用 RT-PCR 技术获得核心片段,结合5′-RACE 和3′-RACE 技术,克隆得到鹰嘴豆生物节律钟基因 LHY 的 cD-NA 全长序列,其核苷酸序列长度为3061 bp,包括2220 bp 的完整开放阅读框(ORF),编码739个氨基酸.验证后命名为 CarLHY 基因,获得基因登录号为 KJ558378.生物信息学研究表明 CarLHY 基因 cDNA 序列与其他植物 LHY 基因具有较高的相似性；预测 CarLHY 蛋白不具有跨膜结构；为转录因子,定位于细胞核中；不具备信号肽.对 CarLHY 蛋白功能结构域预测表明,蛋白质核心结构存在符合转录因子与 DNA 结合的常见功能域 HTH.蛋白系统进化树显示,与大豆分子进化距离最近,其次是黑杨、拟南芥.
종응취두중극륭생물절률종 LHY 기인적 cDNA 전장서렬,진행서렬신식학분석.통과동원극륭책략,이용 RT-PCR 기술획득핵심편단,결합5′-RACE 화3′-RACE 기술,극륭득도응취두생물절률종기인 LHY 적 cD-NA 전장서렬,기핵감산서렬장도위3061 bp,포괄2220 bp 적완정개방열독광(ORF),편마739개안기산.험증후명명위 CarLHY 기인,획득기인등록호위 KJ558378.생물신식학연구표명 CarLHY 기인 cDNA 서렬여기타식물 LHY 기인구유교고적상사성；예측 CarLHY 단백불구유과막결구；위전록인자,정위우세포핵중；불구비신호태.대 CarLHY 단백공능결구역예측표명,단백질핵심결구존재부합전록인자여 DNA 결합적상견공능역 HTH.단백계통진화수현시,여대두분자진화거리최근,기차시흑양、의남개.
The sequence analysis was carried out to investigate the full-length cDNA sequence of biothythm clock gene LHY cloned from Cicer arietinum Linn with 5′-RACE and 3′-RACE.RT-PCR tech-nology was used to obtain core fragment with the homologous cloning strategy.The cDNA sequence was 3061 bp,and the open reading frame (ORF)was 2 220 bp,encoded 739 amino acids.It was named CarLHY after checked.The registration number was KJ558378 in Genebank datebase.The research of biological in-formation science indicated that cDNA sequence of CarLHY gene and other plant LHY gene have higher similarity.It was predicted that CarLHY protein have no transmembrane structure.It was speculated that CarLHY protein have no transmembrane spiral;It was a transcriptional factors and located in the nucleus. It has no signal peptide.CarLHY protein functional domains forecast indicated that there were transcription factor and HTH domain binded to DNA in protein core structure.The phylogenetic tree showed that CarL-HY and Glycine LHY are closest in molecular evolution distance,followed by Populus LHY and Arabii-dopsis thalana LHY gene.