‘鸭梨’钙调素基因的克隆与表达分析

Cloning and expression analysis of calmodulin gene from 'Yali' pears

以‘鸭梨’为材料,用RT-PCR及RACE技术克隆CaM,并借助实时定量PCR技术分析‘鸭梨’花、叶及果实不同发育时期的表达模式,获得了2个长度为764和801bp的CaM cDNA序列,分别命名为PbCaM1和PbCaM2,二者均含有450bp完整开放阅读框,编码149个氨基酸,且氨基酸序列相同。‘鸭梨’CaM在系统进化上具有高度保守性,与苹果相比,两者氨基酸序列为100%一致性。PbCaM1和PbCaM2具有时空表达特异性,它们在成叶、盛花期花瓣和幼果期表达量较高。本研究表明CaM具有高度保守性,PbCaM1和PbCaM2在‘鸭梨’果实早期生长发育过程中起着非常重要的作用。

The calmodulin（CaM）cDNA sequence of‘Yali＇pears was cloned by the technology of RT-PCR and RACE,and the expression patterns of the CaM in flowers,leaves and fruits were analyzed by real-time quantitative PCR.Two CaM cDNA sequences with 764 bp and 801 bp lengths were obtained,being termed as PbCaM1 and PbCaM2 respectively,and both of them contained the intact open reading frame of 450 bp,encoding an identical polypeptide of 149 amino acids.CaM of‘Yali＇pears was highly conservative in phyletic evolution,and exhibited 100%identity to apple CaM across the whole amino acid residues.Further study indicated that PbCaM1 and PbCaM2 genes were temporally and spatially expressed,and showed higher expression levels in petals at bloom stage,mature leaves and young fruits.This study indicated that the CaM was a highly conservative protein,and PbCaM1 and PbCaM2 played important roles in the early development of‘Yali＇pear fruits.