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轮状病毒(rotavirus,RV)非结构蛋白1(non structural protein1,NSP1)在病毒与宿主的相互作用中发挥着重要的功能。运用基因克隆和表达技术在大肠杆菌中表达了TB-Chen株RVNSP1蛋白,进行了NSP1的免疫学性质和RV感染细胞中NSP1蛋白的合成与分布以及NSP1的系统进化和基因分型研究。结果表明,大肠杆菌BL21(DE3)能高效表达重组NSP1蛋白(rNSP1),rNSP1表达量约占菌体总蛋白的34.4%。rNSP1能诱导免疫豚鼠产生特异性血清抗体。Western blot及免疫荧光检测结果表明,抗rNSP1血清抗体能特异性识别自身蛋白,对SA11、Wa株的NSP1蛋白有交叉反应性;免疫荧光结果还表明,SA11感染的MA104细胞中合成的NSP1蛋白在细胞质中区域化聚集形成辐射状排列的颗粒状结构,而Wa株的NSP1不能形成此样结构。至今发现的A组RV至少可以分为16个不同的NSP1基因型,TB-Chen株NSP1为A2型。不同基因型有独特的敏感宿主范围,同一基因型可能感染不同种动物,同一种动物也可能感染不同基因型。基因型A4型和A16型仅在鸟类病毒株中出现;而且鸟类中只有A4型和A16型。研究结果为进一步研究NSP1蛋白质的结构功能及其应用开发奠定了很好的基础。
Rotavirus (RV) Non structural protein 1 (NSP1) plays an important role in the interaction of virus and host. TB-Chen strain RVNSP1 protein was expressed in Escherichia coli by gene cloning and expression technology. The immunological properties of NSP1 and the synthesis and distribution of NSP1 protein in RV infected cells were also studied. The phylogenetic and genotypic studies of NSP1 were also carried out. The results showed that E. coli BL21 (DE3) could efficiently express recombinant NSP1 protein (rNSP1), which accounted for 34.4% of the total bacterial proteins. rNSP1 can induce immune guinea pigs produce specific serum antibodies. The results of Western blot and immunofluorescence showed that the anti-rNSP1 antibody could specifically recognize the self-protein and had cross-reactivity to NSP1 protein of SA11 and Wa strains. The results of immunofluorescence also showed that the NSP1 protein in SA11-infected MA104 cells Regionalization of the cytoplasm to form a radial arrangement of granular structure, and Wa strains of NSP1 can not form such a structure. The group A RVs so far found can be divided into at least 16 different NSP1 genotypes and the TB-Chen strain NSP1 is type A2. Different genotypes have unique sensitive host range, the same genotype may infect different species of animals, the same animal may also be infected with different genotypes. Genotypes A4 and A16 are present only in the avian virus strain; and only birds in the A4 and A16 types. The results laid a good foundation for further study on the structural function of NSP1 protein and its application and development.