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埃博拉病毒(Ebola virus,EBOV)是一种能导致人类及脊椎动物出血热的致死性病毒,对公共卫生具有较严重的危害。EBOV的NP蛋白在病毒复制中具有重要作用,也是诊断该病重要的靶蛋白。文中原核表达重组扎伊尔型EBOV的NP蛋白,重组蛋白免疫bal/c小鼠,制备了一株小鼠抗EBOV-NP的单克隆抗体。利用Western blotting方法,该抗体能特异识别真核表达和原核表达的重组EBOV-NP,并能同莱斯顿型(RestonEbola virus,REBOV)、科特迪瓦型(Cote-d’Ivoire Ebola virus,CIEBOV)和本迪布焦型(Bundibugyo Ebola virus,BEBOV)埃博拉病毒产生交叉反应,而不与苏丹型(the Sudan Ebola virus,SEBOV)和马堡型(the Marburgvirus,MARV)埃博拉病毒产生反应。利用突变PCR和Western blotting方法,定位了该抗体识别的抗原决定簇序列,该序列(PPLESD)位于EBOV-NP蛋白的C端583-588aa。生物信息学研究表明,该序列在已经公布的ZEBOV、CIEBOV、BEBOV共16个型和REBOV的4个型中高度保守。研究结果为建立以上各型埃博拉病毒的检测方法提供了工具,也为研究埃博拉病毒复制及致病机理提供了基础。
Ebola virus (EBOV), a lethal virus that causes hemorrhagic fever in humans and vertebrates, poses more serious public health risks. The NP protein of EBOV plays an important role in viral replication and is also an important target protein for the diagnosis of the disease. In this paper, the NP protein of recombinant Zaire EBOV was expressed in prokaryotic cells and the recombinant protein was used to immunize BAL / c mice to prepare a mouse anti-EBOV-NP monoclonal antibody. Western blotting showed that the recombinant eukaryotic expression vector could specifically recognize eukaryotic and prokaryotic recombinant EBOV-NP and could react with Reston Ebola virus (REBOV), Cote-d’Ivoire Ebola virus (CIEBOV) and The Ebola virus of the Bundibugyo Ebola virus (BEBOV) is cross-reactive and does not react with the Ebola virus of the Sudan (SEBOV) and the Marburg virus (MARV). The mutated PCR and Western blotting methods were used to locate the antigenic determinant sequence of the antibody. This sequence (PPLESD) was located at the C-terminal 583-588aa of EBOV-NP protein. Bioinformatics studies have shown that this sequence is highly conserved among the four published ZEBOV, CIEBOV, BEBOV and 16 REBOV types. The results provide a tool to establish the detection methods of Ebola virus of the above types, and provide a basis for studying the mechanism of Ebola virus replication and pathogenesis.