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CIPK(calcineurin B-like-interacting protein kinase)是一类丝氨酸/苏氨酸蛋白激酶,在植物响应逆境胁迫和激素信号转导中发挥重要作用。本研究利用大豆基因组数据库,在全基因组水平鉴定获得52个CIPK蛋白激酶。蛋白比对分析发现所有Gm CIPK含有高度保守特征性的N端激酶区、连接区和C端调控区。系统进化树分析发现大豆Gm CIPK与拟南芥、水稻CIPK分类一致,分为4个亚家族,且每个亚家族含有3个不同物种的成员,表明Gm CIPK基因的分化早于物种的分化。启动子分析表明,多数Gm CIPK基因的启动子区含有逆境和激素应答元件。组织表达分析发现,Gm CIPK基因呈现多样化的组织表达特性。进一步选取组织表达量相对较高的14个Gm CIPK进行荧光定量PCR分析,结果表明这些菜用大豆CIPK基因在不同程度上均受高温、干旱、高盐胁迫以及ABA、ACC、SA、Me JA激素的诱导表达。采用蛋白同源比对和蛋白互作在线数据库对拟南芥及大豆同源CIPK蛋白激酶与其他蛋白的互作关系进行了预测分析,发现17对同源CIPK与其他蛋白(激酶、磷酸酶、转录因子等)存在互作。本研究为菜用大豆CIPK基因的功能研究与利用奠定了基础。
CIPK is a serine / threonine protein kinase that plays an important role in plant response to stress and hormone signaling. In this study, 52 CIPK protein kinases were identified at the genome level using soybean genome database. Protein alignment analysis revealed that all Gm CIPKs contain highly conserved N-terminal kinase, junction and C-terminal regulatory regions. Phylogenetic analysis showed that Gm CIPK was identical to Arabidopsis thaliana and CIPK, and divided into four subfamilies. Each subfamily contained three members of different species, indicating that Gm CIPK gene differentiation preceded species differentiation. Promoter analysis showed that most Gm CIPK gene promoter regions contain stress and hormone response elements. Tissue expression analysis found that Gm CIPK gene showed a variety of tissue expression characteristics. Fourteen Gm CIPKs with relatively high tissue expression levels were selected for quantitative PCR analysis. The results showed that the CIPK genes in these vegetables were affected by high temperature, drought and salt stress, and ABA, ACC, SA, Me JA hormones to varying degrees Induced expression. The interaction between CIPK protein kinase and other proteins in Arabidopsis thaliana and soybean was predicted by on-line database of protein homology alignment and protein interaction, and 17 pairs of homologous CIPK and other proteins (kinases, phosphatases, Transcription factor, etc.) exist interactions. This study laid the foundation for the functional study and utilization of CIPK gene in vegetable soybean.