摘要
为获得芝麻细胞色素氧化酶Ⅶa亚基基因及其蛋白质序列特征,研究其组织表达特性以及可能的功能。本研究利用NCBI数据库中注释的芝麻COXⅦa基因序列,设计特异PCR引物,以RT-PCR技术进行克隆和表达分析,并利用生物信息学方法分析其序列特征,预测其蛋白质亚细胞定位、跨膜结构和三级结构等。结果表明,克隆的芝麻SiCOXⅦa1和SiCOXⅦa2基因的cDNA序列长度分别为481 bp和416 bp,二者编码的蛋白质序列长度为67个氨基酸残基,具有88%的相似性,属于同源蛋白,均定位于线粒体中,在蛋白序列中35~57 aa形成一个跨膜螺旋区;预测的SiCOXⅦa三级结构均由2段α-螺旋结构组成,但SiCOXⅦa2与SiCOXⅦa1序列的C端形成的高级结构存在较小的差异。表达分析表明,二者在芝麻花蕾、花冠、根、茎和叶中均有表达,且表达模式相似,均在生长旺盛的组织中高丰度表达。本研究为揭示芝麻COX蛋白复合物的结构和功能提供了理论指导。
To obtain cDNA and its protein sequence characteristic of the cytochrome c oxidase 7 a subunits(COX Ⅶa) in Sesamum indicum., and study their tissue expression characteristics and possible functions, the cDNA sequences of COX Ⅶa in Sesamum indicum were annotated in NCBI database. Specific primers were designed for RT-PCR cloning and expression analysis, and the sequence characteristics were analyzed by bioinformatics to predict protein subcellular localization, transmembrane structure and tertiary structure. The results showed that the cloned SiCOX Ⅶa1 and SiCOX Ⅶa2 were 481 bp and 416 bp in length, respectively. The length of the protein sequence encoded by the two genes was 67 amino acid residues with 88% similarity. They belonged to homologous proteins, both located in mitochondria and formed a transmembrane helix region in 35~57 aa of the protein sequence. The predicted tertiary structure of SiCOX Ⅶa1 was composed of two segments of a-helix structure, but there was little difference between the advanced structure formed at C-teminal of SiCOX Ⅶa2 and SiCOX Ⅶa1.The gene expression analysis indicated that SiCOX Ⅶ a1 and SiCOX Ⅶ a2 were expressed in the buds, corolla,roots, stems and leaves of Sesamum indicum, and their expression patterns were similar, and they were abundantly expressed in the growing tissues. This study could provide theoretical guidance for revealing the structure and function of Sesamum indicum COX protein complex.
作者
范小芳
王俊生
刘红占
李伟峰
王瑞霞
胡利宗
高树广
Fan Xiao fang;Wang Junsheng;Liu Hongzhan;Li Weifeng;Wang Ruixia;Hu Lizong;Gao Shuguang(College of Life Sciences and Agriculture, Zhoukou Normal University, Zhoukou, 466001;Zhoukou Academy of Agriculture Sciences, Zhoukou, 466001)
出处
《分子植物育种》
CAS
CSCD
北大核心
2019年第12期3825-3833,共9页
Molecular Plant Breeding
基金
国家特色油料产业技术体系项目(CARS-14-2-17)
河南省教育厅重点攻关项目(18A210006)共同资助
