玉米锌指蛋白基因ZmRFP的克隆及表达特性

Cloning and Expression Characteristics of Zinc Finger Protein Gene ZmRFP from Maize(Zea mays L.)

锌指蛋白(ZFP)是一类重要的转录因子,通过与核酸相互作用来调控基因的表达,在植物生长发育与衰老以及非生物胁迫等方面具有重要作用。本研究从玉米自交系‘辽3162’中克隆了一个C3H2C3型锌指蛋白基因ZmRFP,该基因开放阅读框长度为1563 bp,编码520个氨基酸,编码蛋白预测分子量为57.06 kD,等电点为6.18,为亲水性蛋白。保守结构域和跨膜结构域分析表明该蛋白存在环锌指蛋白超家族RING_Ubox结构域,属于锌指蛋白家族转录因子,不存在跨膜结构域,为非分泌蛋白。对该蛋白进行二级结构和三级结构预测,氨基酸残基主要以无规则卷曲为主。对其顺式作用元件进行分析发现,其上游2000 bp内存在多个与非生物胁迫和植物激素相关的作用元件。进化树分析表明,RFP蛋白的进化高度保守,与高粱的进化关系最为相近。器官特异性分析发现,该基因在玉米叶片中的表达量最高,在盐和干旱胁迫下的表达研究发现,该基因对非生物胁迫存在响应。ZmRFP基因的克隆和表达分析为进一步研究其生物学功能和应用提供参考。

Zinc finger protein(ZFP)is a kind of important transcription factor,which regulates gene expression by interacting with nucleic acid,it plays an important role in plant growth,senescence and abiotic stress.In this study,we cloned a C3H2C3 zinc finger protein gene(ZmRFP)from maize inbred line'Liao3162'.The open reading frame length of ZmRFP was 1563 bp,encoding a protein containing 520 amino acids.The predicted molecular weight was 57.06 kD,and the isoelectric point was 6.18.Analysis of conservative domain and transmembrane domain showed that the protein belonged to zinc finger protein family transcription factor with RING domain,it was also an non-secretory transmembrane domain protein.The amino acid residues were mainly random CRIMP based on prediction of the secondary structure and the tertiary structure.There were several components related to abiotic stress and plant hormones in the upstream 2000 bp.Phylogenetic analysis showed that the evolution of RFP proteins was highly conservative and most similar to that of sorghum.Organ specific analysis showed that the expression of this gene was the highest in maize leaves.Under abiotic stress,the gene expression was induced under salinity and drought stress.Cloning and expression analysis of ZmRFP gene in maize laid a foundation for further study of its biological function and application.