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采用SMART-RACE-PCR技术获得了茶树细胞周期蛋白依赖激酶基因(CsCDK1)的全长cDNA序列,并进行了相关的生物信息学分析。采用实时定量PCR研究了该基因在茶树越冬芽休眠到萌发后不同阶段的表达模式。克隆到茶树CsCDK1cDNA序列1245bp(GenBank登录号JF449383),其开放阅读框长924bp,编码307个氨基酸,预测分子量为34.52kD,具有CDKs家族典型的保守结构域和空间结构,属于B型CDK家族。系统进化分析表明,CsCDK1氨基酸序列与猕猴桃的CDK亲缘关系最近,达到96%,与其它植物的CDK序列相似性亦在80%以上。该基因在茶树越冬芽休眠期的表达量低于恢复生长期,在萌发期表达量最高。说明该基因与茶树越冬芽休眠的解除关系密切。
The full-length cDNA sequence of cyclin-dependent kinase gene (CsCDK1) in tea plant was obtained by SMART-RACE-PCR and bioinformatics analysis was carried out. Real-time quantitative PCR was used to study the expression patterns of this gene at different stages after dormancy until germination. The cDNA sequence of CsCDK1 was cloned into GenBank accession number JF449383, which contained 924 bp open reading frame (ORF) encoding 307 amino acids with a predicted molecular mass of 34.52 kD. It has a conserved CDKs family structure and spatial structure and belongs to the B-type CDK family. Phylogenetic analysis showed that the amino acid sequence of CsCDK1 was closest to CDK in kiwifruit, reached 96%, and similar to other plant CDK sequences in more than 80%. The expression level of this gene in the dormant period of the overwintering buds of tea plants was lower than that of the restoring growth phase, and the expression level was the highest during the germination phase. This gene is more closely related to the release of the dormancy of the overwintering buds of tea trees.