牡丹DNA甲基结合域蛋白基因PsMBD5的克隆和表达特性分析

发布时间：2018-01-20 22:57

本文关键词： PsMBD RACE 生物信息学分析表达特性　出处：《华北农学报》2017年02期 　论文类型：期刊论文

【摘要】：前期通过转录组结合表达谱分析发现一类MBD片段在低温解除牡丹花芽内休眠中的差异表达,为了进一步研究MBD基因特征及其在牡丹休眠解除中的作用,利用RACE扩增获得1204bpPsMBD5全长cDNA,其中编码框1056bp,推测编码351个氨基酸。生物信息学分析结果显示,PsMBD5蛋白分子量约为38.1274kDa,等电点为5.14,不稳定系数为44.27,推测该蛋白为不稳定蛋白。同源性分析表明,牡丹PsMBD5与梅PmMBD5的同源性最高,为38.1%,与亚麻荠CsMBD5的同源性最低,为25.8%,与13种拟南芥MBD蛋白同源性最高的是AtMBD5。实时荧光定量PCR分析表明,PsMBD5基因在牡丹不同组织中均有表达,在初花期牡丹的根中转录水平最高,在花瓣、心皮和萼片中次之,在雄蕊中最低;PsMBD5基因在牡丹花芽中受低温诱导,且转录水平在低温处理14d时达到最大值,之后维持较高的转录水平。研究结果为进一步解析PsMBD5基因对牡丹花芽的休眠解除的调控机制奠定基础。
[Abstract]:In order to further study the characteristics of MBD gene and its role in dormancy release of peony, the differential expression of a class of MBD fragments was found in the early stage by transcriptome binding expression analysis in order to further study the characteristics of MBD gene and its role in dormancy release of peony buds at low temperature. The full-length cDNAof 1204bpPsMBD5 was obtained by RACE amplification, and the encoding frame 1056bpwas deduced to encode 351 amino acids. The molecular weight of PsMBD5 protein was about 38.1274 kDa, the isoelectric point was 5.14, and the coefficient of instability was 44.27. The homology between PsMBD5 and PmMBD5 was the highest (38. 1), and the homology with CsMBD5 was 25.8%. The highest homology with 13 Arabidopsis thaliana MBD proteins was AtMBD5.The real-time fluorescence quantitative PCR analysis showed that the PsMBD5 gene was expressed in different tissues of Paeonia suffruticosa. The transcription level was the highest in the roots of peony at the beginning of flowering, followed by petal, carpels and sepals, and the lowest in stamens. The PsMBD5 gene was induced by low temperature in the bud of Peony, and the transcription level reached the maximum at 14 days after low temperature treatment. The results laid a foundation for further analysis of the regulation mechanism of PsMBD5 gene on dormancy release of peony bud.
【作者单位】：青岛农业大学生命科学学院山东省高校植物生物技术重点实验室;青岛农业大学农学与植保学院;
【基金】：国家自然基金面上项目(31471908;31372104)
【分类号】：S685.11
【正文快照】： DNA甲基化现象是DNA主要的一种表观遗传修饰,在植物的生长发育、花芽分化、非生物胁迫、基因组防御、基因表达调控等方面发挥着重要作用[1]。DNA甲基结合域(Methyl-binding domain,MBD)蛋白是一类转录因子,与甲基化DNA结合有关,在DNA甲基化依赖的基因调控过程中起作用[2]。近

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