【摘要】 旨在分别构建NA蛋白和M2蛋白的重组质粒,并进行表达鉴定。采用PCR扩增了H9N2分离株A/chicken/Shandong/LY1/2017毒株的NA基因和M2基因。同源性分析证实,这2个基因序列与目前流行毒株的基因同源性很高。将NA基因克隆至pcold1原核表达载体,将M2基因克隆至pET28a原核表达载体,成功构建了pcold1-NA和pET28a-M2重组质粒,并成功表达出相应的融合蛋白。然后对表达的融合蛋白进行纯化,并采用免疫印迹验证其免疫反应性。本试验为进一步研究H9N2亚型禽流感病毒致病机制及检测技术奠定了基础。更多还原

【Abstract】 The purpose of this study was to construct recombinant plasmids of NA and M2 proteins and identify their expression. The NA and M2 genes of the H9N2 strains A/chicken/Shandong/LY1/2017 were amplified by PCR, respectively. Homology analysis was performed and the result confirmed that the two gene sequences had high homology with that of the current epidemic H9N2 strains. The NA gene was cloned into the prokaryotic expression vector pcold1 and the M2 gene was cloned into the prokaryotic expression vector pET28a. Recombinant plasmids of pcold1-NA and pET28a-M2 were successfully constructed and the corresponding fusion proteins were successfully expressed in BL21, respectively. Then, the expressed fusion proteins were purified and their immunogenicity was verified by Western blotting. The sequence analysis and expressions of these two genes laid a good foundation for further research on pathogenesis and diagnostic techniques of the avian influenza H9N2 subtype.更多还原