甘蔗细胞色素C基因的电子克隆与分析

Electronic cloning and characterization of sugarcane ScCyt C gene using bioinformatics tools

以甘蔗类似细胞色素C的EST序列CF576943.1为探针,通过电子克隆技术获得了甘蔗细胞色素C基因(Cytochrome C,Cyt C)的一条cDNA全长序列,命名为ScCyt C。用生物信息学方法对该基因氨基酸序列与组成、亚细胞定位、跨膜区与信号肽、疏水性/亲水性、蛋白质二、三级结构以及功能等进行分析与预测。结果表明:Cyt C基因全长1 073 bp,编码112个氨基酸,该基因位于细胞质,为非分泌型蛋白,无规卷曲为主要二级结构原件,含有1个保守功能域,主要功能为翻译并且在不同植物中具有高度保守性。电子表达分析结果显示,该基因在甘蔗各个组织均有表达,其中在茎和根中的表达量比其他组织类型中表达量高,此外,该基因的表达可能受到低温的调控。为甘蔗细胞色素C基因的结构及其功能的研究奠定了一定的基础。

The full-length cDNA sequence of one sugarcane cytochrome C gene （ScCyt C gene） was obtained by in silico cloning using CF576943.1 sequence from Saccharum sinense as the probe sequence. Some characters of the ScCyt C gene encoding amino acid, including the composition of amino acid sequence, physical and chemical properties, subcellular localization, transmembrane domain, hydrophobicity / hydrophilicity, secondary and tertiary structure of protein plus functional domains, were analyzed by bioinformatics tools. The results showed that the full-length ScCyt C gene from sugarcane is 1073 bp which encodes a polypeptide of 112 amino acids. The encoded protein of ScCyt C gene, with a conserved domain sequences, is soluble and located in cytoplasm or chloroplast, and the corresponding secondary structure of this protein is mainly composed of random coil. The function of the ScCyt C protein is mainly involved with translation. It is highly conservative in different plants. Electronic expression analysis revealed that the ScCyt C gene is constitutively expressed in all the types of sugarcane tissues, and the expression of this gene in the stem and root of sugarcane is much higher than that in all the other types of sugarcane tissues. In addition, the expression of this gene is regulated under the stress of low temperature. The results obtained in this study will provide reference for molecular cloning, structural analysis and function identification of ScCyt C gene in sugarcane.