摘要
以大豆“Williams 82”为试验材料,利用RT-PCR法从中克隆得到转录因子GmTGA15基因。生物信息学分析表明,GmTGA15基因组序列包括8个外显子和7个内含子,开放阅读框为1 089 bp,编码362个氨基酸;蛋白理化性质分析表明,GmTGA15蛋白分子量为41.01 kDa,理论等电点为6.28,为亲水性蛋白;蛋白一级结构具有1个典型的bZIP保守结构域,属于bZIP转录因子家族,与其他15种植物TGA(TGACG motif-binding factor)蛋白同源比对及系统进化分析显示,均具有相同的bZIP保守结构域,与同科植物野大豆亲缘关系最近,为97.25%;其蛋白二级结构预测α-螺旋为主,占63.81%。通过酵母单杂试验验证GmTGA15具有转录激活活性;利用PEG4000介导法将融合表达载体转化拟南芥原生质体,发现GmTGA15定位于细胞核。TGA转录因子在植物抵御逆境胁迫及生长发育中起重要作用。此结果将为GmTGA15基因功能的进一步研究提供基础,也为植物抗逆调控机制的研究提供理论参考。
GmTGA15 gene was cloned by reverse transcription PCR(RT-PCR)from soybean“Williams 82”.Bioinformatics analysis shows that the genome sequence of GmTGA15 includes 8 exons and 7 introns and GmTGA15 contains a complete open reading frame(ORF)of 1089 bp encoding of 362 amino acid residues.Analysis of the physicochemical properties of the GmTGA15 protein shows that the molecular mass was 41.01 kDa,and the theoretical isoelectric point is 6.28,which is a hydrophilic protein;The primary structure of the GmTGA15 protein has a typical bZIP conserved domain,belonging to the family of bZIP transcription factors.Homologous alignments and phylogenetic analysis of amino acid sequence show that GmTGA15 protein and TGA proteins of 15 other plants all have the same bZIP conserved domain.GmTGA15 protein is most closely related to wild soybean GsTGA,and the consistency of sequence alignment is 97.25%.Secondary structure prediction of GmTGA15 shows thatα-helices are dominated,accounting for 63.81%.GmTGA15 possessed transcriptional activation activity by the method of a yeast one-hybrid assay.The fusion expression vector was introduced into Arabidopsis protoplasts by a PEG4000-mediated method and the results showed that GmTGA15 was localized in the nucleus.TGA transcription factors play an important role in plants'resistance to stress and growth.The results of this study can provide basis for further exploration of the function of GmTGA15 gene.Meanwhile,it can provide theoretical reference for study on plant stressresistant regulation mechanism in the future.
作者
袁学顺
周佳玉
刘淼
周莹
姚丹
崔喜艳
陈展宇
YUAN Xueshun;ZHOU Jiayu;LIU Miao;ZHOU Ying;YAO Dan;CUI Xiyan;CHEN Zhanyu(College of Life Sciences,Jilin Agricultural University,Changchun 130118,China;College of Agronomy,Jilin Agricultural University,Changchun 130118,China)
出处
《吉林农业大学学报》
CAS
CSCD
北大核心
2024年第3期358-365,共8页
Journal of Jilin Agricultural University
基金
吉林省自然科学基金项目(20210101023JC,20220101337JC)。
关键词
大豆
TGA转录因子
转录激活活性
亚细胞定位
soybean
TGA transcription factor
transcriptional activation activity
subcellular localization
