大豆GmNAC46生物信息学、转录自激活及组织表达分析

Bioinformatics,Autoactivation,and Tissue Expression Analysis of Soybean GmNAC46

NAC转录因子是一类广泛存在于植物中且为植物所特有的转录因子超家族,不仅调控植物的生长发育等方面,同时也响应植物受到的生物及非生物胁迫。为了研究大豆中NAC转录因子的功能,利用PCR的方法从大豆中克隆GmNAC46基因,采用生物信息学方法对该基因及编码的蛋白质进行分析,采用酵母单杂交实验分析转录因子的自激活活性,采用qRT-PCR的方法分析基因的组织特异性表达。结果表明:GmNAC46基因开放阅读框长1218 bp,编码405个氨基酸,蛋白质分子量为45.872 kD,等电点为4.88。GmNAC46蛋白在4~156位氨基酸处含有1个高度保守的NAC结构域,属于亲水性蛋白,定位于细胞核中。GmNAC46蛋白与野生大豆的NAC蛋白亲缘关系较近。酵母单杂交实验结果表明,转录因子GmNAC46具有转录自激活活性。组织特异性表达模式分析表明,GmNAC46基因在大豆根、茎、叶、子叶中都表达,在根中表达量最高。

NAC transcription factors are a superfamily of transcription factors that exist widely in plants and are unique to plants.They not only regulate plant growth and development,but also respond to biological and abiotic stresses.In order to study the function of NAC transcription factors in soybean,the GmNAC46 gene was cloned from soybean by PCR,bioinformatics method was used to analyze the gene and its encoded protein,the autoactivation activity of transcription factors was analyzed by yeast single hybrid experiment.The tissue-specific expression of the gene was analyzed by qRT-PCR.The results showed that:The open reading frame was 1218 bp,and encoding 405 amino acids,the molecular weight of the protein was 45.872 kD,and the isoelectric point was 4.88.GmNAC46 protein contained a highly conserved NAC domain at amino acids 4-156,which was a hydrophilic protein and was localized in the nucleus.GmNAC46 protein was closely related to NAC protein of wild soybean.The results of yeast single hybridization experiment showed that GmNAC46 had transcriptional activation activity.Tissue specific expression pattern analysis showed that GmNAC46 was expressed in root,stem,leaf and cotyledon,and the highest expression level was found in roots.