呼吸道合胞病毒融合蛋白F1和截短F1蛋白的表达差异研究  

The research on different expression between F1 protein and truncated F1 protein of respiratory syncytial virus

作　　者：傅生芳[1] 安静[1] 朱传凤[1] 寇桂英[1] 陈汉泉[1] 余黎[1] 周旭[1] 

机构地区：[1]兰州生物制品研究所,兰州730046

出　　处：《微生物学免疫学进展》2012年第1期10-14,共5页Progress In Microbiology and Immunology

摘　　要：目的构建呼吸道合胞病毒融合蛋白F1和截短F1蛋白的原核表达载体,并对它们在大肠杆菌中的表达差异进行了初步研究。方法用DNAstar软件对呼吸道合胞病毒F1蛋白进行亲疏水性和抗原表位可能性分析后,将其两端的疏水区域截去之后与pET-42b(+)构建表达载体,同时用相同的表达系统构建F1蛋白的表达载体并将2种重组蛋白进行诱导表达。实验对2种蛋白在Rossata/pET-42b(+)菌株中的表达难易度、表达形式及初步洗涤的包涵体纯度进行了比较。结果与F1蛋白相比,截短的F1蛋白相对更容易表达,表达的可溶性蛋白含量更高,洗涤纯化后的包涵体纯度也更高。结论呼吸道合胞病毒F1蛋白截去两端疏水氨基酸后更容易表达,为后期蛋白的大量制备及其免疫原性研究奠定了基础。Objective To construct the prokaryotic expression vectors of F1 protein and truncated F1 protein of human respiratory syncytial virus and induce expression,the difference between them was primarily researched. Methods The hydrophilicity and antigen epitop of F1 protein of human respiratory syncytial virus were analyzed with DNAstar software,then the hydrobility regions of F1 subunit were truncated.The truncated F1 and F1 was inserted into pET-42b（＋） vector respectively,then they were induced to express in Rossata strain.The expressed degree,expressed form and crude purified inclusion body of them were compared and analyzed.Results The results suggested that the truncated F1 was more easily express than F1,and truncated that of F1 had higher soluble target protein content and higher purity of inclusion body.Conclusion F1 protein can be more easily expressed after truncat the hydrophobic regions of C-and N-terminus,which laid a foundation of preparation and immunogenicity research of this antigen.

关 键 词：呼吸道合胞病毒 F1 截短F1 表达差异 

分 类 号：R373[医药卫生—病原生物学]