Cloning and Bioinformatics Analysis of ZmMAPKKK21 Gene in Maize
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丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)在植物应对生物和非生物胁迫过程中发挥着重要作用.在前期研究中我们挖掘到ZmMAPKKK21这一潜在的重要抗旱基因,本研究从玉米抗旱自交系J24中克隆获得ZmMAPKKK21基因,并对其进行生物信息学分析.结果表明,玉米ZmMAPKKK21基因开放阅读框长1419bp,编码472个氦基酸,是不具有信号肽的亲水性蛋白,且其启动子序列含有多个与逆境胁迫和激素有关的顺式作用元件;玉米中的MAPKKK21蛋白预测定位在细胞核上,蛋白互作预测结果显示,ZmMAPKKK21蛋白与参与植物逆境胁迫相关的MAPKK3蛋白和ZIM家族蛋白发生互作,其结构与高粱(Sorghum bicolorL.)和谷子(Setaria italica L.)等高抗旱禾本科作物相比,不但在STKc_MAPKKK结构域上高度保守,且具有更加相似的二级结构和三级结构;qRT-PCR分析发现,ZmMAPKKK21基因在玉米根系中表达水平较高,干旱胁迫后该基因在根和叶中表达上调,进一步验证了 ZmMAPKKK21基因的表达与干旱胁迫响应的相关性.
The mitogen-activated protein kinase(MAPK)plays an important role in the process of plants responsing to both biological and non-biological stresses.In the previous study,we excavated the potentially important drought-resistant gene ZmMAPKKK21.In this study,the ZmMAPKKK21 gene was cloned from the drought-resistant maize inbred line J24 and bioinformatics analysis was performed on the gene.The findings indicated that the ZmMAPKKK21 gene's open reading frame measured 1419 bp and encoded 472 amino acids.The protein was hydrophilic and devoid of a signal peptide.Its promoter sequence featured several cis-acting regions associated with hormones and stress.It is anticipated that the maize MAPKKK21 protein was found on the nucleus.The outcome of the protein interaction prediction indicated that ZmMAPKKK21 interacted with the plant stress-related proteins MAPKK3 and ZIM family.In addition to being extremely conservative on the STKc_MAPKKK domain,its secondary and tertiary structures are more akin to those of highly drought-resistant gramineous crops,such as sorghum(Sorghum bicolor L.)and foxtail millet(Setaria italica L.).The association between the expression of the ZmMAPKKK21 gene and the response to drought stress was further confirmed by qRT-PCR analysis,which revealed that the gene's expression level was high in maize roots and that it was up-regulated in roots and leaves following drought stress.
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