日本结缕草ZjHY5的克隆、亚细胞定位及转录自激活检测

Cloning,Subcellular Localization and Autonomous Transcriptional Activation Testing of Transcription Factors ZjHY5 in Zoysia japonica

转录因子LONG HYPOCOTYL 5(HY5)在光诱导基因表达中起中心调控因子的作用。为了探究日本结缕草(Zoysia japonica)中ZjHY5基因的亚细胞定位及自激活特性,本研究从日本结缕草(Zoysia japonica)中克隆得到ZjHY5基因。该基因开放阅读框为711 bp,编码236个氨基酸残基。ZjHY5蛋白无信号肽,进化树分析显示其与漆姑草(Sagina japonica)及苜蓿(Medicago sativa)中HY5蛋白的相似度最高。亚细胞定位结果显示,ZjHY5定位于细胞核中。通过构建酵母诱饵载体,并对其进行自激活检测,结果显示,ZjHY5基因无自激活活性,可用于后续试验。本试验为深入研究日本结缕草ZjHY5基因的功能奠定了基础。

The transcription factor LONG HYPOCOTYL 5(HY5)acts as a central regulator in light-induced gene expression.To investigate the subcellular localization and self-activation characteristics of ZjHY5 gene in Zoysia japonica,ZjHY5 gene was cloned.The open reading frame of this gene is 711 bp,which encode 236 amino acids.ZjHY5 protein has no signal peptide,and phylogenetic tree analysis showed that it had the highest similarity to HY5 protein in Sagina japonica and Medicago sativa.The results of subcellular localization showed that ZjHY5 resided in the nucleus.The yeast bait vector was constructed to detect the self-activation of ZjHY5 protein.The results showed that ZjHY5 gene had no self-activation activity,which could be used in subsequent yeast two hybrid experiments.This study laid a foundation for further study on the function of ZjHY5 gene in Zoysia japonica.