草莓钙依赖蛋白激酶基因的克隆与表达分析  

Gene Cloning and Expression Analysis of Calcium-dependent Protein Kinase in Strawberry

作　　者：王萌[1,2,3] 王俊[1] 刘洪[2,3] 李静[2,3] 段可[2] 叶正文[3] 高清华[3] 

机构地区：[1]宁夏大学生命科学学院,银川750021 [2]上海市农业科学院生物技术研究所,上海201106 [3]上海市农业科学院林木果树研究所,上海市设施园艺技术重点实验室,上海201106

出　　处：《西北植物学报》2011年第5期875-880,共6页Acta Botanica Boreali-Occidentalia Sinica

基　　金：上海农科院科技发展基金[农科发2008(02)];上海市科技兴农重点攻关项目[沪农科攻字(2008)第1-4号];上海市科委自然科学基金(10ZR1426700),上海市科委青年科技启明星计划(09QA1405300)

摘　　要：以EST测序结合RACE技术,从久香草莓的茎尖中分离得到一个新基因,BLASTX分析显示这是一个钙依赖蛋白激酶编码基因,与GenBank中的FaCDPK1同源,命名为FaCDPK2,GenBank登录号为HQ829293.采用半定量RT-PCR对FaCDPK2基因在不同组织中的表达分析表明,FaCDPK2全长3 775 bp,读码框1 659bp,由12个外显子组成,编码552个氨基酸残基,在第89-349位是保守的蛋白激酶结构域,C端含4个EFhand钙结合域.该基因编码蛋白与拟南芥CPK28序列一致性为86.25%,与草莓FaCDPK1序列一致性为43.99%.表达分析显示该基因表达范围较广,在所分析的七种草莓组织材料中,以花中表达水平最高,在果实中的表达可能受成熟的诱导.EST sequencing project combined with the RACE（rapid amplification of cDNA ends） technique led to the isolation of a novel gene from shoot apical meristems of Fragaria×ananassa Duch.cv.Jiuxiang.BLAST analysis revealed that this gene is homologous to FaCDPK1 in Genbank,thus it was named FaCDPK2 and submitted to Genbank under the Accession No.HQ829293.Semi-quantitative RT-PCR was further performed to study its expression pattern in different tissues.The genomic sequence for FaCDPK2 is 3 775 bp in length,consisting of 12 exons.This gene harbors a 1 659 bp open reading frame（ORF） encoding a 552 amino acid peptide,with a protein kinase domain（amino acids 89 to 349） in the middle region and four EFhand calcium-binding sites at the C terminal.The deduced amino acid sequence shows 86.25% identity to CPK28 from Arabidopsis thaliana,while only 43.99% identity to FaCDPK1 from strawberry.Semi-quantitative RT-PCR analysis showed that this gene was expressed in all tissues tested,but the expression level was the highest in flower and probably induced by ripening process in fruit.

关 键 词：久香草莓 CDPK基因 克隆 基因结构 进化树 RT-PCR 

分 类 号：Q789[生物学—分子生物学]