信号肽生物信息学分析在Neurospora crassa phyA基因鉴定中的应用

Application of bioinformatics analysis of signal peptide in the identification of Neurospora crassa phyA gene

目的对Neurospora crassa基因组中与黑曲霉phyA具有46.85%相似性的EAA33149.1蛋白基因进行鉴定并确定其功能。方法利用信号肽预测软件SignalP 3.0 Server,真核蛋白序列中精氨酸和赖氨酸前导肽切割位点以及信号肽切割位点预测软件ProP 1.0 server,跨膜结构域预测软件Tmpred和TMHMM Server v.2.0,GP I锚定位点预测软件big-P I Predictor以及亚细胞器中蛋白定位分布预测软件TargetP v1.01对EAA33149.1蛋白基因进行预测分析,根据分析结果将该基因克隆到酿酒酵母中表达。结果确定了该蛋白的信号肽、信号肽切割位点以及分泌途径。重组酿酒酵母表达的54000重组蛋白,显示了植酸酶活性。结论初步确定该蛋白基因为N.crassa phyA基因。

Objective To identify the function of the gene encoding Neurospora crassa EAA33149.1 protein which has 46.85% similarity with Aspergillus niger phA gene. Methods The bioinformatics analysis was conducted using the prediction algorithms SignalP v3.0, arginine and lysine propeptide cleavage sites in eukaryotic protein sequence prediction algorithms ProP 1.0 server, transmembrane domain prediction algorithms Tmpred and TMHMM v2.0, potential GP I-anchor sites prediction algorithm big-P I Predictor and the subcellular protein location prediction algorithms TargetP v1.01. According to the analysis results, the gene was cloned into Saccharomyces cerevisiae. Results The signal peptide, the cleavage site and the secretion pathway were determined, and the expressed recombinant protein with 54 000 displayed phytase activity. Conclusion The gene has been identified to encode N. crassa phyA.