香石竹Dccdc20基因的克隆及表达分析

Cloning and Expression Analysis of Dccdc20 Gene from Carnation

减数分裂是香石竹遗传改良的重要环节,Cdc20蛋白在减数分裂中起着重要作用。为了更好的了解Cdc20在香石竹减数分裂中的作用机制。本研究利用RT-qPCR结合RACE技术,从香石竹花药中分离克隆了一个Cdc20基因的全长c DNA序列,命名为Dccdc20 (GenBanK登录号:2217177)。Dccdc20基因全长1 847 bp,含有439个氨基酸,长1 320 bp的开放阅读框。蛋白序列比对显示Dccdc20基因与其他物种的Cdc20基因有很高的相似度,RT-qPCR结果显示Dccdc20基因的表达量随着花药发育呈现递减趋势,在花蕾长度为1.1~1.2 cm时期的花药中表达最高,而在叶和茎中表达量较低,研究表明该基因可能在调节香石竹减数分裂中发挥作用。

Meiosis is an important part of genetic improvement of carnation.Cdc20 protein plays an important role in meiosis.In order to better understand the mechanism of Cdc20 in carnations meiosis.In this study,a fulllength cDNA sequence of Cdc20 gene was isolated and cloned from anther of carnation by RT-qPCR and RACE technology and named as Dccdc20.The full length c DNA sequence of Dccdc20 is 1847 bp,with an ORF of 1320 bp encoded 439 amino acids.The alignment of the protein sequence showed that the Dccdc20 gene hasa high similarity with the Cdc20 of other species.Real time qRT-PCR showed that the expression level of the Dccdc20,with the development of meiosis,was obviously decreased,and got to the maximum at anthers of 1.1~1.2 cm length flower bud.While the expression in the leaves and stems were lower.Studies have speculated that this gene may play a role in regulating carnation meiosis.