论文部分内容阅读
用RT PCR方法扩增流感病毒血凝素基因 ,将其克隆到 5型腺病毒载体质粒 pAd5C中。用此质粒与EcoRI酶切回收的 5型腺病毒基因组大片段共转染 2 93细胞 ,通过PCR初步筛选得到重组病毒。SDS PAGE电泳、West ernblot证明重组病毒能表达流感病毒的血凝素蛋白。此重组病毒经滴鼻免疫Balb/C小鼠 ,ELISA实验证明能诱导小鼠产生针对流感病毒的循环抗体IgG和分泌型抗体IgA。本实验证明 ,利用E3区缺失的 5型腺病毒载体可以用来表达流感病毒血凝素抗原 ,重组抗原具有良好的免疫原性 ,有可能发展成为基因工程流感疫苗
The influenza virus hemagglutinin gene was amplified by RT PCR and cloned into adenovirus type 5 adenovirus plasmid pAd5C. A total of 293 cells were co-transfected with the large fragment of adenovirus type 5 adenovirus isolated by this plasmid and EcoRI, and the recombinant virus was screened by PCR. SDS PAGE electrophoresis, West ernblot proves that the recombinant virus can express influenza virus hemagglutinin protein. The recombinant virus was intranasally immunized Balb / C mice, ELISA experiments proved that mice can be induced to produce circulating antibodies against influenza virus IgG and secretory antibody IgA. This experiment proved that the use of deletion of the E3 region type 5 adenovirus vector can be used to express influenza virus hemagglutinin antigen, recombinant antigen has good immunogenicity, it may develop into a genetically engineered influenza vaccine