甘蔗CDK基因的cDNA全长克隆与表达分析

发布时间：2018-06-29 00:44

本文选题：甘蔗 + CDK基因　；参考：《作物学报》2017年01期

【摘要】：植物细胞周期依赖性蛋白激酶(cyclin-dependent kinases,CDKs)是一类丝氨酸/苏氨酸蛋白激酶,与周期蛋白(Cyclins)协同作用,是重要的细胞周期性调控因子。本研究从甘蔗受花叶病毒侵染的转录组数据库中获得一条与高粱(Sorghum bicolor)CDK基因(Gen Bank登录号为XP_002466536.1)高度同源的Unigene序列,通过RT-PCR扩增获得长度为1799 bp的甘蔗Sc CDK基因(Gen Bank登录号为KR258796)。序列分析显示Sc CDK基因含一个完整的开放阅读框(65~1603 bp),编码512个氨基酸,且具有CDK典型保守结构域,如ATP结合位点、磷酸结合位点及活化环A-loop。此外,生物信息学预测显示该基因编码的蛋白定位于细胞核,为可溶性蛋白,不存在信号肽,二级结构元件多为无规则卷曲,主要参与蛋白翻译。实时荧光定量PCR分析表明,Sc CDK基因的表达具有组织特异性,其在蔗芽上的表达量最高,其次是在蔗肉、蔗根、叶鞘和蔗皮中;在PEG、Na Cl和ABA的胁迫诱导过程中,Sc CDK均表现上调作用,且受ABA胁迫后表达量最高,约为对照组的1.9倍。推测该基因的表达与甘蔗抗干旱和抗渗透胁迫有关,同时受ABA的诱导,参与细胞周期分裂。
[Abstract]:Plant cell cycle dependent protein kinases (cyclin-dependent kinases) are a kind of serine / threonine protein kinases, which interact with cyclins and are important cell cycle regulators. In this study, a highly homologous Unigene sequence of Sorghum bicolor CDK gene (GenBank accession number XP002466536.1) was obtained from the transcriptional database of sugarcane infected by Mosaic virus, and the length of SC CDK gene (GenBank accession number was KR258796) was obtained by RT-PCR amplification. Sequence analysis showed that Sc CDK gene contained a complete open reading frame (65m1603 bp), encoding 512 amino acids) and had typical conserved domains of CDK, such as ATP-binding sites, phosphoric acid binding sites and activation ring A-loops. In addition, bioinformatics prediction showed that the protein encoded by the gene was located in the nucleus and was soluble protein, and there was no signal peptide. The secondary structural elements were mostly irregular curl, mainly involved in protein translation. Real-time fluorescence quantitative PCR analysis showed that the expression of Sc CDK gene was tissue specific, and the highest expression was found in sugarcane bud, followed by sugarcane meat, root, leaf sheath and bark. During the stress induction of PEGN NaCl and ABA, Sc CDK was up-regulated, and the highest expression level was about 1.9 times as much as that of the control group under ABA stress. It is inferred that the expression of this gene is related to resistance to drought and osmotic stress in sugarcane and is induced by ABA to participate in cell cycle division.
【作者单位】：福建农林大学/农业部福建甘蔗生物学与遗传育种重点实验室/国家甘蔗产业技术研发中心;
【基金】：福建省杰出青年科学基金(2015J06006) 福建省高等学校新世纪优秀人才支持计划(JA14095) 引进国际先进农业科学技术计划(948计划)项目(2014-S18) 国家现代农业产业技术体系建设专项(CARS-20)项目资助~~
【分类号】：S566.1

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