本研究采用RT-PCR技术,克隆获得禾谷镰刀菌(Fusarium graminearum)致病性CYP51C蛋白基因c DNA序列,并对蛋白氨基酸序列进行生物信息学分析,明确其序列典型特征。结果表明:其c DNA序列全长为1 554 bp,编码517个氨基酸;该蛋白分子量为58.6 k D,等电点为6.36;含有细胞色素P450家族成员典型的保守结构域;不具有信号肽,属于非分泌性蛋白;具有跨膜结构域,是亲水性蛋白;蛋白质二级结构最主要的结构元件是α螺旋和无规则卷曲;亚细胞定位预测显示CYP51C蛋白主要位于细胞质中。这些研究结果为蛋白纯化及其蛋白结构生物学研究提供参考,进而可为病原真菌药物靶标设计奠定基础。
In this study,the full -length cDNA sequence of pathogenic protein CYP51C from Fusarium graminearum was cloned using RT -PCR,and the bioinformatics analysis of amino acid sequences was con-ducted and its typical characteristics was identified.The results showed that the full -length cDNA sequence of CYP51C included a complete open reading frame (ORF)of 1 554 bp,which encoded 517 amino acids.Its molecular weight was 58.6 kD and its isoelectric point was 6.36.It possessed the conserved domains which were characteristics of cytochrome P450 family members.It was not a secretory protein without a signal pep-tide,but was a hydrophilicity protein with transmembrane domain.The major structural elements of polypep-tide chain were α-helix and random coil.Subcellular localization result indicated that CYP51C was mainly located in cytoplasm.These results would provide references for the study on protein purification and protein structural biology,and then lay a foundation for drug design targeting pathogenic fungi.