日本结缕草ZjTCP4基因的克隆、转录自激活检测及表达模式分析

Cloning,Autonomous Transcriptional Activation Detection and Expression Pattern Analysis of Zj TCP4 Gene in Zoysia japonica

TCP家族蛋白是植物中一种特有的转录因子,在植物的发育过程中起着重要的作用。本研究从日本结缕草(Zoysia japonica)中克隆ZjTCP4基因(GenBank登录号:MZ540773),该基因完整编码区为2097 bp,编码699个氨基酸。进化树及保守序列分析显示,ZjTCP4蛋白含有TCP家族高度保守的螺旋-环-螺旋的TCP结构域,与玉米(Zea mays)中的ZmTCP4蛋白同源关系最高,且具有相似的保守基序构成,属于TCP转录因子家族中的CIN亚类。酵母自激活检测结果显示,ZjTCP4不具备自激活活性,可进行后续酵母双杂交实验及互做蛋白检测分析。ZjTCP4基因表达模式分析显示,其在结缕草衰老叶片及根系中的表达量较高。对ZjTCP4蛋白的烟草(Nicotiana tabacum)亚细胞定位实验显示,该蛋白定位于细胞质内。本研究为进一步研究日本结缕草ZjTCP4蛋白及TCP转录因子家族提供了科学依据。

TCP family proteins are unique transcription factors in plants and play an important role in plant development.The ZjTCP4 gene was cloned from Zoysia japonica in this study(GenBank accession number:MZ540773).The complete coding region of the gene was 2097 bp,encoding 699 amino acids.The analysis of evolutionary tree and conserved sequence showed that ZjTCP4 protein contained TCP domain of highly conserved helix-loop-helix in TCP family,which had the highest homology with ZmTCP4 protein in Zea mays and had similar conserved motif composition.ZjTCP4 belongs to the CIN subclass of the TCP transcrition factor family.According to the results of yeast self-activation,the ZjTCP4 does not have self-activation activity,so it can be used for yeast two-hybrid experiments and protein detection and analysis.The expression pattern analysis of the ZjTCP4 gene showed that it was highly expressed in aged leaves and roots of Zoysia.The subcellular localization of ZjTCP4 protein in Nicotiana tabacum indicated that the protein was localized in the cytoplasm.This study provides a scientific basis for further study of Zoysia japonica ZjTCP4 protein and TCP transcription factor family.