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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cDNA全长序列,其核苷酸序列长度为3 061 bp,包括2 220 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。
The cDNA sequence of LHY gene was cloned from chick pea and sequenced. The cDNA fragment of LHY gene was cloned by homology cloning strategy using RT-PCR and 5’-RACE and 3’-RACE techniques. The length of nucleotide sequence of LHY was 3 061 bp, including 2 220 bp complete open reading frame (ORF), encoding 739 amino acids. After validation, the gene was named CarLHY gene, accession number is KJ558378. Bioinformatics studies show that CarLHY gene cDNA sequence and other plant LHY gene has a high similarity; CarLHY protein is not predicted transmembrane structure; as a transcription factor, located in the nucleus; do not have the signal peptide. Prediction of the CarLHY protein functional domain shows that there is a common functional domain HTH in the protein core structure which is compatible with the transcription factor and DNA binding. Phylogenetic tree of phylogenetic tree showed that the closest evolutionary distance to soybean was followed by Populus nigra and Arabidopsis thaliana.