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.
出处
《西北植物学报》
CAS
CSCD
北大核心
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)
关键词
久香草莓
CDPK基因
克隆
基因结构
进化树
RT-PCR
Fragaria×ananassa Duch.cv.Jiuxiang
gene coding calcium-dependent protein kinase
cloning
gene organization
phylogenetic tree
RT-PCR
