菜心Dof基因克隆、生物信息学分析及表达分析

Cloning,Bioinformatics Analysis and Expression Analysis of Dof from Flowering Chinese Cabbage

Dof (DNA binding with one finger)是植物特有的转录因子,在调控植物生长发育和代谢过程中发挥着重要作用。为明确Dof具体的生物学功能,本研究以‘油绿501’菜心为材料,采用同源克隆法获得了Dof全长序列,并对其编码蛋白结构、基因表达模式及其互作蛋白的调控网络进行分析。结果表明:菜心BcDof3;1基因全长624 bp,编码207个氨基酸,其N-端含有保守的Dof锌指结构。系统进化树分析表明,BcDof3;1与白菜BrDof3;1和拟南芥AtDof2的同源关系较近。RT-qPCR结果表明,BcDof3;1在菜心根、茎、叶、花和种子中均有表达,尤其在叶和花中的表达水平最高;其表达水平受氮调控。互作蛋白网络分析表明,BcDof3;1可能通过中性转化酶、淀粉酶、酰胺酶等调控碳氮代谢。研究结果为深入研究Dof在菜心生长发育和碳氮代谢中提供了理论参考。

As a kind of plant-specific transcription factor,Dofs(DNA binding with one finger) play important roles in regulating plant growth,development and metabolism.To understand the biological functions of Dof in plant,a full-length cDNA of Dof was cloned by homologous cloning method from flowering Chinese cabbage(Brassica campestris L.ssp.chinensis var.utilis Tsen et Lee) variety ’Youlv501’.Meanwhile,the bioinformatics analysis of BcDof3;1 protein and its regulatory network of interacting proteins were analyzed by using the online software,and the expression of BcDof3;1 was detected by quantitative real-time PCR(RT-qPCR).The results showed that the sequence of BcDof3;1 was 624 bp,encoding 207 amino acid residues,N terminal of which contained a conserved motif of zinc finger.Phylogenetic analysis showed that BcDof3;1 was antigenically closely related to BrDof3;1 from Brassica rapa and At Dof2 from Arabidopsis thaliana.The results of RT-qPCR showed that BcDof3;1 expressed in root,stem,leaf,flower and seed of B.campestris,especially in leaf or flower,the ones of which was higher;moreover,its expression level was affected by the starvation or adding of N.The interactional network indicated that BcDof3;1 may regulate the metabolism of carbon and nitrogen by neutral invertase,amylase or amylase.The results of this study will provide a theoretical reference for further study the function of Dof in the growth and development and the metabolism of carbon and nitrogen in B.campestris.