基于生物信息学方法预测筛选Blag2的B细胞和T细胞表位及突变体-细胞生物学专业论文.docxVIP

万方数据 万方数据 【关键词】 德国小蠊；Bla g 2；B、T细胞表位；氨基酸突变；分子动力学 模拟 III In Silico Prediction and Screening of the Mutants, B and T-cell Epitopes of Bla g 2 Major: Cell Biology Postgraduate: Yang Tie Supervisor: Zhang Chunlin [Abstract] Objective: In Silico Prediction and Screening the B and T-cell Epitopes of Bla g 2 . To obtain the key amino acids by evaluating the site-directed binding energy, and then validate it by molecular dynamics, so as to provide new evidence for the key amino acids on producing the hypoallergenic derivative and allergen vaccine of Bla g 2. Methods: ① Three immunoinformatics tools, the DNAStar Protean system, the Bioinformatics Predicted Antigenic Peptides system (BPAP) and the BepiPred 1.0 server, were used to predict the potential B-cell epitopes, while Net-MHCIIpan-2.0 and NetMHCII-2.2 were used to predict the T-cell epitopes of Bla g 2. ②To obtain the key amino acid sites of the mutation through screening and evaluation with the FoldX software. ③The wild type complex(wcBla g 2) and the mutanted complexs (Bla g 2-D201A ,Bla g 2-E233A,Bla g 2-E233R,Bla g 2-K251A) were built by Swiss-PDBviewer software. ④ The molecular dynamics simulations of the five built compounds were made with GROMACS software, and the fluctuation of interactive influence between the molecular binding energy with the complex before and after mutation and the wild type and 4 mutated complexes was computed. Results: ① Eight B cell epitopes of Bla g 2 were predicted, namely 55KYEKLKP61,79SAVGRGIE86,124AAPGCPNAL132,157RFQDGE162,200GDTTVAPAG TQAIID214,223PKAYVNPINE233,241TRRICKLDCSKIPSL255,288PXGHSDHF295. In the T-cell prediction, the Bla g 2 allergen was predicted to have two strongly binding nonamer core epitope sequences (IC5050 nm) and 34 weakly binding sequences (50 nmIC50500 nm). ②Seven key amino acid sties of mutants were identified through fold X software, which were 200th,201th,202th amino acid sites in “200GDTTVAPAGTQAIID214; 230th,233th amino