忍冬ICE1基因的cDNA克隆及表达分析

Cloning and expression analysis of ICE1 gene in Lonicera japonica

采用RT-PCR技术克隆得到忍冬(Lonicera japonica)ICE1(inducer of CBF expression 1)基因LjICE1(GenBank:MK584895),其cDNA序列全长为1575 bp,编码524个氨基酸,理论等电点5.58,为不稳定亲水性蛋白,含多个糖基化和磷酸化位点,且具有类似MYC元件的bHLH结构。同源比对发现ICE1蛋白含有富S区、NLS区、bHLH结构域、Zipper结构域和跨膜区,系统进化分析发现LjICE1与茶树(Camellia sinensis)的亲缘关系最近。RT-qPCR结果表明,LjICE1基因在低温胁迫3 h时表达量最高。本研究为进一步解析忍冬低温响应机制提供了基因资源。

The ICE1(inducer of CBF expression)gene LjICE1 was cloned by RT-PCR technology from Lonicera japonica,the full-length cDNA of LjICE1 gene was 1575 bp,encoding 524 amino acids with a theoretical isoelectric point of 5.58.It is an unstable hydrophilic protein with multiple glycosylation and phosphorylation sites and has bHLH structure is similar to MYC.The homologous alignment revealed that the ICE1 protein contained the S-rich region,the NLS region,the bHLH domain,the Zipper domain and the transmembrane region.The phylogenetic analysis found that LjICE1 was closely related to the Camellia sinensis.The results of RT-qPCR showed that the expression of LjICE1 gene was highest at 3 h under low temperature stress.This may provide gene resources for further analysis of the low temperature response mechanism of Lonicera japonica.