甘蔗细胞色素P450还原酶基因的电子克隆与分析

Electronic cloning and characterization of sugarcane ScCYP450 gene using bioinformatics tools

旨在为甘蔗细胞色素P450还原酶基因的实验克隆、功能鉴定及其应用提供参考。本研究以甘蔗类似细胞色素P450还原酶基因的EST序列CF576130.1为探针,在甘蔗EST数据库中进行检索,并基于电子克隆技术获得了甘蔗细胞色素P450还原酶基因(Cytochrome P450 reductase)的一条cDNA全长序列,命名为ScCYP450。采用生物信息学方法,对该基因编码蛋白从氨基酸组成、理化性质、亚细胞定位、跨膜结构域、疏水性/亲水性、高级结构及功能域等方面进行预测和分析。结果表明该基因全长1 821 bp,包含一个744 bp的开放阅读框,编码247个氨基酸,该基因编码蛋白定位于细胞内质网(膜),为可溶性碱性蛋白,不存在信号肽,二级结构原件多为无规卷曲,含有多个保守功能域,主要功能为辅酶因子生物合成和翻译功能。

The present study aims to provide references for research on the experimental cloning, functionidentification and its application of sugarcane Cytochrome P450 reductase gene. The full-length cDNA sequence ofone sugarcane Cytochrome P450 reductase gene termed ScCYP450, was obtained by in silico cloning usingCF576130.1 sequence from sugarcane as the probe sequence. Using bioinformatics methods, several characteristicsof ScCYP450 encoding amino acids, including the amino acid composition, physical and chemical properties, thesubcellular localization, the transmembrane domain structure, hydrophobic/hydrophilic, advanced structure andforecast and plus the functional domains, were analyzed. The result showed that this gene is 1 821 bp length andcontains a 744 bp open reading frame, encoding 247 amino acids, and the encoded protein localizes in cellsendoplasmic reticulum（film）, as the alkaline soluble protein. There is no signal peptide. The secondary structuremostly consistes of random coil. ScCYP450 protein contains several conservative functional domains, and the mainfunction of this protein involved in growth factor biosynthesis and transcription.