烟草NtCIPK2基因的克隆及表达分析

发布时间：2018-07-03 15:39

本文选题：烟草 + NtCIPK　；参考：《浙江农业学报》2017年10期

【摘要】：CIPK(CBL-interacting protein kinase)是一类具有生物活性的丝/苏氨酸蛋白激酶,通过与类钙调神经素B亚基蛋白(Calcineurin B-like protein,CBL)相互作用从而调节植物在适应或抵制非生物逆境胁迫中的生理活动。基于同源序列克隆烟草K326中Nt CIPK2基因c DNA全长,采用q RT-PCR分析Nt CIPK2的组织及逆境胁迫下特异性表达。结果表明,该基因全长1 341 bp,编码446个氨基酸残基,预测分子量为50.3 ku,等电点为8.90。同源性分析结果显示,该蛋白与绒毛状烟草(Nicotiana tomentosiformis)CIPK2有95%的同源性,故命名为Nt CIPK2。生物信息学分析表明,该蛋白是一个亲水蛋白,其N端含有活化环结构域S_TKc,C端含有调节结构域CIPK-C,可能定位在质膜。组织表达分析发现,该基因在烟草根、茎、叶、花中均有表达,其中茎中表达量最高。逆境胁迫试验表明,该基因受低钾、高盐、干旱、H2O2和ABA诱导,参与烟草非生物逆境胁迫的响应。
[Abstract]:Cipk (CBL-interacting protein kinase) is a kind of biologically active silk / threonine protein kinase, which regulates the physiological activities of plants in adapting to or resisting abiotic stress by interacting with Calcineurin B subunit protein (CBL). The full-length cDNA of Nt-CIPK2 gene was cloned from tobacco K326 based on homologous sequence, and the specific expression of Nt-CIPK2 in tissue and under stress was analyzed by qRT-PCR. The results showed that the total length of the gene was 1 341 BP, encoding 446 amino acid residues, the predicted molecular weight was 50.3 kuand and the isoelectric point was 8.90. The result of homology analysis showed that the protein had 95% homology with Nicotiana tomentosiformis CIPK2, so it was named NT CIPK2. Bioinformatics analysis showed that the protein was a hydrophilic protein, and the N-terminal of the protein contained the activated ring domain (SSP), which contained the regulatory domain CIPK-C, which may be located in the plasma membrane. Tissue expression analysis showed that the gene was expressed in the roots, stems, leaves and flowers of tobacco, and the highest expression was found in the stems. The stress test showed that the gene was induced by low potassium, high salt, H2O2 and ABA, and was involved in the response to abiotic stress in tobacco.
【作者单位】：四川农业大学农学院;
【基金】：植物生理学与生物化学国家重点实验室开放课题(SKLPPBKF 1505,SKLPPBKF 1506)
【分类号】：Q943.2;S572

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