烟草NtCIPK2基因的克隆及表达分析

doi:10.3969/j.issn.1004-1524.2017.10.01

浙江农业学报 ›› 2017, Vol. 29 ›› Issue (10): 1597-1604.DOI: 10.3969/j.issn.1004-1524.2017.10.01

烟草NtCIPK2基因的克隆及表达分析

卓维, 陈倩, 鲁黎明, 李立芹^*

四川农业大学农学院,四川成都 611130

收稿日期:2017-04-14 出版日期:2017-10-20 发布日期:2017-12-05
通讯作者: 李立芹,E-mail:liliqin88@163.com
作者简介:卓维(1994-),女,重庆人,硕士研究生,主要从事植物分子生物学研究。E-mail: 992167898@qq.com
基金资助:
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)

Cloning and expression analysis of NtCIPK2 gene in Nicotiana tabacum

ZHUO Wei, CHEN Qian, LU Liming, LI Liqin^*

College of Agronomy, Sichuan Agricultural University, Chengdu 611130, China

Received:2017-04-14 Online:2017-10-20 Published:2017-12-05

摘要/Abstract

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NtCIPK2基因cDNA全长,采用qRT-PCR分析NtCIPK2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NtCIPK2。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。

关键词: 烟草, NtCIPK2, 克隆, 生物信息学, 基因表达

Abstract: CIPK(CBL-interacting protein kinase)is a class of bioactive serine/ threonine protein kinase that interacts with the calcineurin B-like protein (CBL) to regulate the physiological activity of plants in adapting or resisting to abiotic stress. The homologous sequences were used to clone the full length cDNA of NtCIPK2 gene in the tobacco cultivar K326, and qRT-PCR was used to analyze the tissue-specific and abiotic stress expression pattern of NtCIPK2. The results showed that this gene contained 1 341 bp and encoded 446 amino acid. The predicted molecular weight was 50.3 ku and the isoelectric point (pI) was 8.90. The homology analysis showed that the protein had 95% homology with Nicotiana tomentosiformis CIPK2, thus it was named as NtCIPK2. Bioinformatics analysis showed that NtCIPK2 was a hydrophilic protein whose N-terminus contained a conserved active loop domain S_TKc and the C-terminus contained the regulatory domain CIPK-C, and might be localized in the plasma membrane. Expression patterns showed that the gene was expressed in roots, stems, leaves and flowers in mature stage, which has the highest expression in stems. Expression patterns under abiotic stress indicated the gene expression was induced by low-potassium, high-salt, drought, H₂O₂ and ABA treatment, and involved in the response to abiotic stress in Nicotiana tabacum.

Key words: tobacco, NtCIPK2, cloning, bioinformatics, gene expression

中图分类号:

S572

卓维, 陈倩, 鲁黎明, 李立芹. 烟草NtCIPK2基因的克隆及表达分析[J]. 浙江农业学报, 2017, 29(10): 1597-1604.

ZHUO Wei, CHEN Qian, LU Liming, LI Liqin. Cloning and expression analysis of NtCIPK2 gene in Nicotiana tabacum[J]. , 2017, 29(10): 1597-1604.

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烟草NtCIPK2基因的克隆及表达分析

Cloning and expression analysis of NtCIPK2 gene in Nicotiana tabacum

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