高羊茅FeTOC1基因的克隆、差异表达及亚细胞定位分析

拟南芥TOC1基因是中央振荡器的重要组成部分,其编码的蛋白质TOC1通过光周期途径调控拟南芥对光照的响应.为了揭示高羊茅FeTOC1基因的生物学功能,本研究通过同源克隆获得该基因全长cDNA序列,对其在不同光照水平下的表达模式进行分析,并对其编码的蛋白质进行亚细胞定位.结果显示,高羊茅FeTOC1基因的全长cDNA序列...

Cloning,Differential Expression and Subcellular Localization of FeTOC1 Gene in Tall Fescue

TOC1 gene is a major component of the central oscillator,which regulates the response to illumination through photoperiod pathway in Arabidopsis thaliana.In order to illustrate the biological function of FeTOC1 in Festuca elata,we obtained the full-length cDNA sequence of FeTOC1 by homologous cloning,analyzed the expression patterns of FeTOC1 under diverse light levels,and conducted the subcellular localization of FeTOC1.The results indicated that the full-length of FeTOC1 was 1557 bp,encoding 517 amino acid residues;the ended protein FeTOC1 contained the pseudo-receiver domain of about 120 amino acids at N-terminus and the short CCT motif of about 50 amino acids at C-terminus;the phylogenetic tree analysis showed that FeTOC1 had close relations to those in barley,wheat and coarse goat grass,which were up to 99%.Then,the constructed target protein was fused with the GFP expression vector,and injected into tobacco,the expression suggested that FeTOC1 was mainly located in the nucleus.Real-time fluorescence quantitative analysis of the expression patterns of FeTOC1 in leaves revealed that the expression of FeTOC1 had a diurnal rhythm under both long-and short-day,and the expression at illumination 0~4 h reached the highest;compared with continuous darkness,continuous illumination weakened the oscillatory rhythm of FeTOC1 dramatically;reversing the circadian rhythm,FeTOC1 was preferentially regulated by environmental signal.To sum up,FeTOC1 gene played an important role in circadian clock regulation in tall fescue.