甘蔗钙依赖蛋白激酶基因克隆与序列分析

发布时间：2018-06-03 13:55

本文选题：甘蔗 + 钙依赖蛋白激酶基因(CDPK)　；参考：《南方农业学报》2017年04期

【摘要】：【目的】克隆甘蔗钙依赖蛋白激酶基因(ScCDPK1),为其功能解析和育种利用打下基础。【方法】根据甘蔗cDNA文库中的CDPK基因序列信息设计引物,利用RT-PCR克隆ScCDPK1基因,并采用在线生物信息学分析软件对其推导蛋白的理化性质、跨膜结构、信号肽及保守结构域等进行预测分析。【结果】克隆获得的ScCDPK1基因完整编码区大小1650 bp,编码549个氨基酸,相对分子量61.971 kD,等电点(pI)6.78,不稳定指数35.63。同源性和遗传进化树分析结果显示,ScCDPK1基因与小麦TaCPK7基因亲缘关系最近,同源性达88%;ScCDPK1蛋白与其他已知CDPK蛋白一致,具有CDPKs家族特有的4个保守区域(可变区、蛋白激酶区、自抑制区和钙离子结合区),其二级结构主要由α螺旋(42.80%)和无规则卷曲(32.97%)构成;蛋白功能分类预测结果显示,ScCDPK1蛋白是一种阳离子通道蛋白,可能参与植物胁迫应答、信号转导和翻译调控等生物反应。【结论】ScCDPK1基因编码的蛋白与其他已知CDPK蛋白一致,具有CDPKs家族特有的4个保守区域,是一种阳离子通道蛋白。
[Abstract]:[objective] to clone SCCDPK1 gene from sugarcane and lay a foundation for its function analysis and breeding. [methods] based on the sequence information of CDPK gene in sugarcane cDNA library, primers were designed and RT-PCR was used to clone ScCDPK1 gene. The physicochemical properties, transmembrane structure, signal peptide and conserved domain of the deduced protein were predicted by on-line bioinformatics analysis software. [results] the complete coding region of ScCDPK1 gene was 1650 BP, encoding 549 amino acids. The relative molecular weight is 61.971 KD, the isoelectric point (Pi) is 6.78, and the instability index is 35.63. The results of homology and phylogenetic tree analysis showed that ScCDPK1 gene was closely related to wheat TaCPK7 gene. The homology of ScCDPK1 protein was consistent with other known CDPK proteins, and had four conserved regions (variable region, protein kinase region) specific to the CDPKs family. The secondary structure of SCPK1 protein was mainly composed of 伪 helix 42.80) and irregular crimp 32.97), and the results of protein functional classification predicted that ScCDPK1 protein was a cationic channel protein, which might participate in plant stress response, and its secondary structure was mainly composed of 伪 helix (42.80) and irregular curl (32.97). [conclusion] the protein encoded by ScCDPK1 gene is consistent with other known CDPK proteins and has four conserved regions of the CDPKs family, which is a cationic channel protein.
【作者单位】：广西作物遗传改良生物技术重点开放实验室;广西农业科学院甘蔗研究所/农业部广西甘蔗生物技术与遗传改良重点实验室;贺州学院;
【基金】：广西自然科学基金项目(2013GXNSFAA019077,2013GXNSFAA019088) 广西八桂学者专项项目([2013]3号) 广西农业科学院科技发展基金项目(2016JM05)
【分类号】：S566.1

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