摘要
为了寻求一种有效的疫苗用于预防小反刍兽疫(PPR),运用RT-PCR方法扩增PPRV的F基因并将其克隆到pCAGGS载体中,构建了真核表达质粒pCAGGS-F。将构建的重组质粒转染哺乳动物细胞上进行瞬时表达,通过Western-blotting、间接免疫荧光检测证实F蛋白得到了高效表达。将重组质粒作为DNA疫苗,通过脚掌皮内注射免疫BALB/c小鼠,采用间接ELISA检测抗体的产生情况。结果显示,pCAGGS-F重组质粒在有无佐剂CpG的情况下均能诱导小鼠产生较高的抗体水平。
To investigate a vaccine against peste des petits ruminants（PPR）,F gene of PPR virus was amplified by RT-PCR and cloned into the eukaryotic expression vector pCAGGS to construct recombinant plasmid pCAGGS-F. Then the constructed plasmid was transfected into BHK-21 and COS7 cells and the expression of F protein could be detected by indirect immunofluorescence assay and Western-blot. Further- more,the immunogenicity was evaluated in BALB/c mice with pCAGGS-F via intracutaneous injection in the foot and the antibody was detected through indirect ELISA. In result, pCAGGS-F was able to induce specific immune responses in mice.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2013年第5期500-504,共5页
Chinese Veterinary Science
基金
现代农业产业技术体系建设专项(CARS-40-10)
甘肃省农业生物技术研究与应用开发项目(GNSW-2010-08)
