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目的用生物信息学方法分析刚地弓形虫抑制蛋白(T.gondii profilin,TgPRF)的主要特性与抗原表位,筛选该蛋白的T/B细胞联合抗原表位。方法利用NCBI上的ORF finder、ProtParam、SignalP、TMHMM、MotifScan、WoLF PSORT、PSORTⅡ、SOPMA、Bcepred、SYFPEITHI、NetCTL等生物信息学在线分析程序,结合Gene Runner和DNAMAN等生物信息学软件,分析并预测TgPRF蛋白的开放阅读框、理化性质、信号肽、跨膜区、翻译后修饰位点、亚细胞定位、二级结构、表面可及性、可塑性、亲(疏)水性及T/B细胞抗原表位。结果 TgPRF基因的开放式阅读框为492个碱基的核苷酸序列。TgPRF蛋白由163个氨基酸组成,相对分子质量为17 555.2,分子式为C_(764)H~(1160)N_(204)O_(258)S_7,等电点为4.40,有4个表面可及性参数≥1.9的区域、5个亲水性参数得分≥1.9的区域、3个柔韧性参数得分≥2的区域、10个翻译后修饰位点、8个潜在B细胞抗原表位和3个T/B细胞联合表位。结论 TgPRF蛋白有多个优势抗原表位,为后续弓形虫免疫保护性的研究提供理论指导。
Objective To analyze the main features and antigenic epitopes of T. gondii profilin (TgPRF) using bioinformatics methods and screen the T / B cell epitopes of the protein. Methods Bioinformatics online analysis programs such as ORF finder, ProtParam, SignalP, TMHMM, MotifScan, WoLF PSORT, PSORTⅡ, SOPMA, Bcepred, SYFPEITHI and NetCTL on NCBI were combined with bioinformatics software such as Gene Runner and DNAMAN to analyze and predict TgPRF open reading frame, physical and chemical properties, signal peptide, transmembrane region, posttranslational modification sites, subcellular localization, secondary structure, surface accessibility, plasticity, affinity Bit. As a result, the open reading frame of the TgPRF gene is a 492-nucleotide nucleotide sequence. The TgPRF protein consists of 163 amino acids and has a relative molecular mass of 17 555.2, a molecular formula of C 764 H 1160 N 204 O 258 258, an isoelectric point of 4.40, and four surface accessibility parameters Five regions with a score of ≥1.9 in the hydrophilic parameters, three regions with a score of ≥2 in the flexibility parameters, ten posttranslational modification sites, eight potential B-cell epitopes and three T / B Cell joint epitope. Conclusion TgPRF protein has multiple dominant epitopes, which may provide theoretical guidance for the subsequent study on the immunoprotection of Toxoplasma gondii.