铁皮石斛DoMAPK5基因的克隆及表达特征分析

发布时间：2018-06-29 12:51

本文选题：铁皮石斛 + 促分裂原蛋白激酶　；参考：《时珍国医国药》2016年02期

【摘要】：目的克隆及分析铁皮石斛DoMAPK5基因的表达特征。方法利用cDNA末端快速克隆技术(RACE),首次从铁皮石斛接菌共生萌发种子中分离得到1个新的MAPK基因,对其编码蛋白的理化性质、保守结构域等特征及进行了分析。并应用荧光定量PCR对DoMAPK5基因在石斛不同组织中表达模式分析。结果 DoMAPK5基因(Gen Bank注册号KJ472788),全长为2184bp、编码一条597个氨基酸的肽链,预测分子量为67.39KDa,等电点为9.28,编码的蛋白均不含信号肽,在210-232位具有23个氨基酸的跨膜域,具有MAPK蛋白家族保守的丝氨酸/苏氨酸蛋白激酶的结构域及MAPK位点。序列比对及系统发育树分析结果表明DoMAPK5基因与葡萄MAPK基因均具有很高的相似性(82%),与水稻MAPK10亲缘关系最近。应用荧光定量PCR对DoMAPK5基因在石斛不同组织中表达模式分析显示,该基因在未接菌的植物组织中属组成型表达,在接菌共生萌发的植物组织中显著上调,为未萌发种子的10.49倍。结论 DoMAPK5在植物组织中属于组成型表达,具有受菌根真菌诱导表达的特征。
[Abstract]:Objective to clone and analyze the expression of DoMAPK5 gene in Dendrobium candidum. Methods A new MAPK gene was isolated from the seeds of Dendrobium candidum by cDNA terminal rapid cloning (race) for the first time. The physicochemical properties, conserved domain and other characteristics of the encoded protein were analyzed. The expression patterns of DoMAPK5 gene in different tissues of Dendrobium candidum were analyzed by fluorescence quantitative PCR. Results DoMAPK5 gene (GenBank registration number KJ472788) was 2184bp. it encodes a peptide chain of 597 amino acids. The predicted molecular weight is 67.39 KDa. the isoelectric point is 9.28, and the encoded protein contains no signal peptide and has 23 amino acid transmembrane domains at 210-232 sites. The domain and MAPK site of serine / threonine protein kinase were conserved by MAPK family. Sequence alignment and phylogenetic tree analysis showed that there was a high similarity between DoMAPK5 gene and grape MAPK gene (82%), and the relationship between DoMAPK5 gene and rice MAPK10 was the closest. The expression patterns of DoMAPK5 gene in different tissues of Dendrobium candidum were analyzed by fluorescence quantitative PCR. The results showed that the expression of DoMAPK5 gene in non-inoculated plant tissues was constitutively expressed, and was significantly up-regulated in inoculated symbiotic germinating plant tissues, 10.49 times as much as that of non-germinated seeds. Conclusion DoMAPK5 is constitutively expressed in plant tissues and is induced by mycorrhizal fungi.
【作者单位】：北华大学医学部;中国医学科学院北京协和医学院药用植物研究所;
【基金】：国家自然科学基金(No.31170314) 国家教育部博士点基金项目(No.20131106130002)
【分类号】：S567.239

【参考文献】