茶树细胞周期蛋白基因的克隆与表达  

Cloning and Expression Analysis of Cyclin Gene(CsCYC1) of Tea Plant

作　　者：王新超[1] 杨亚军[1] 马春雷[1] 金基强[1] 马建强[1] 曹红利[1] 

机构地区：[1]中国农业科学院茶叶研究所/国家茶树改良中心,杭州310008

出　　处：《西北植物学报》2011年第12期2365-2372,共8页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家自然科学基金项目(30872059);现代农业(茶叶)产业技术体系项目(CARS-23)

摘　　要：以茶树萌动芽为材料,采用SMART-RACE-PCR技术从茶树萌动芽中获得了茶树细胞周期蛋白基因的全长cDNA序列(命名为CsCYC1),并用实时定量PCR方法(qRT-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。结果显示:(1)该基因全长1 956bp,包含1 320bp的开放阅读框(ORF),编码439个氨基酸残基。(2)CsCYC1预测分子量为49.35kD,具有细胞周期蛋白家族典型的保守cyclin-box结构域和三维结构。(3)系统进化分析结果表明,CsCYC1的氨基酸序列与葡萄、蓖麻、毛果杨、琴叶鼠耳芥、拟南芥等的相似性分别为77%、74%、72%、68%和67%。(4)实时荧光定量PCR分析显示,CsCYC1基因在茶树越冬芽休眠期的表达量远低于恢复生长期,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 eDNA of eyclin gene （CsCYC1） was cloned from the sprouting buds of tea plant （Camellia sinensis） by SMART-RACE-PCR based on the partial sequences in sprouting bud SSH-cDNA li- brary. The sequence and structure characteristics of its encoded protein were analyzed, and its expression patterns during the time of bud dormancy and bud break were evaluated by quantitative real-time PCR （qRT-PCR）. The full length eDNA is 1956 bp （GenBank Accession No. JF795471）,which includes 1 320 bp ORF and encodes 439 amino acid residues with a putative molecular mass of 49.35 kD. CsCYC1 has a typical conserved cyclin-box domain near the C-terminus and 3D structure of cyclin family. Phylogenetic a- nalysis showed that CsCYC1 had 77%,74%,72%,68% and 67% similarity with cyclin of Vitis vinifera, Ricinus communis , Populus tomentosa ,Arabidopsis l yrata and Arabidopsis thaliana , respectively. The re- sults of quantitative real-time PCR showed that the expression level of CsCYC1 at dormant stages was low- er than that at growth stages, and the highest expression level emerged at sprouting stage. It indicated that CsCYC1 had very close relationship with tea plant budbreak in sprinm

关 键 词：茶树 芽萌发 细胞周期蛋白基因CsCYC1 克隆 实时定量PCR 

分 类 号：Q786[生物学—分子生物学]