融合表达猪源C3d与猪流感病毒HA DNA疫苗的构建

为了构建融合表达分子佐剂猪补体C3d与猪流感病毒血凝素(HA)的真核表达载体,论文克隆了猪C3d全长基因与H3N2亚型猪流感病毒的HA基因,并构建了包含3拷贝的C3d和经过改造的HA(替换信号肽并去除跨膜区)的质粒pHA-C3d3.序列分析表明,获得的猪C3d与参考序列相比核苷酸同源性达到99.7%,氨基酸同源性达99...

Construction of DNA Vaccines Expressing Fusion of C3d-HA Against Swine Influenza Virus

The pig C3d gene and HA gene of swine influenza virus were amplified by PCR in order to construct DNA vaccines expressing fusion of C3d-HA against swine influenza virus.Homology comparison between the sequence of pig C3d in this study and those of reference pig C3d showed 99.7% identities at nucleotide level,and 99.3% identities at amino acid level.The amino acid sequence of CR2 binding domain of C3d in this study was identical with reference sequence.Homology comparison between the sequence of pig C3d in this study and those of C3d from mouse,bovine,goat,human and chicken showed 80% identities and 62.5% respectively.The CR2 binding domain of pig C3d(p28) has 4-7 and 17 amino acid differences compared with those of mouse,bovine,goat,human and chicken.This suggested that the C3d of mammal and avian C3d had obviously different.293T cells were transfected by the plasmid pHA-C3d3 with lipofectamine 2000.Western blot assay showed that the fusion C3d-HA was detected in supernatants and cell lysates.This study laid a good foundation to further research a novel DNA vaccine against swine influenza virus.