华支睾吸虫多药物/代谢产物外排蛋白样基因的识别和生物信息学分析

目的预测华支睾吸虫假想的多药物/代谢产物外排蛋白的结构及功能及其应用前景。方法利用多种生物信息学分析软件,从华支睾吸虫全长cDNA质粒文库中识别出多药物/代谢产物外排蛋白样基因,分析蛋白的拓扑学结构、生物学和免疫学功能特征,根据结构与功能预测其应用前景。结果该基因编码189个氨基酸;含有5段跨膜区,N端在胞内,C端在胞外,具有主要协助因子超家族的结构特征。与日本血吸虫同源蛋白(一致性为67%,相似性为80%)相比,该蛋白的4段跨膜区中含有保守的精氨酸和组氨酸残基。该蛋白含有多个磷酸化位点和T、B细胞表位,且线性B细胞表位均位于胞外区。结论所克隆的华支睾吸虫基因可能编码一个小蛋白型阴离子多药物/代谢物外排蛋白;该蛋白可能定位于华支睾吸虫皮层表膜,是一个潜在的诊断和疫苗候选抗原和药物靶标分子。

Recognization and bioinformatics analysis of Clonorchis sinensis small multidrug/metabolites efflux protein-like gene

A clone from Clonorchis sinensis full-length cDNA plasmid library was recognized as a putative small multidrug/metabolites efflux protein-like gene,and the structural and functional characteristics of the putative protein were analyzed by on-line bioinformatics tools provided by NCBI(http://www.ncbi.nlm.nih.gov/,ExPaSy(http://www.ncbi.nlm.nih.gov/),and CBS Prediction Servers(http://www.cbs.dtu.dk/services/) and Vector NTI suite 8.0 software package,and its application was predicted on the basis of the bioinformatics.It was found that gene contained a complete cds encoding 189 amino acids containing 5 transmembrane regions with the N terminus inside of the cytoplasm membrane and C terminus outside.It belong to the subfamily of MFS-1 of the Major Facility Superfamily with the property of small multidrug/metabolites efflux protein,which was characterized by 4 or 5 transmembrane regions.Comparing with the homologue of Schistosoma japonicum(with identity of 67% and similarity of 80%),each of four of five transmembrane regions contains one or two conserved basic amino acid residue arginine or histidine and there are several phosphorylation sites and linear T cell and B cell epitopes mainly located in the outside of membrane.These suggest the protein maybe locate in the membrane of tegument of C.sinensis and is involved in the pump-out of anionic drugs or deleterious metabolites,and is a potential candidate of vaccine and immunodiagnosis for clonorchiasis.