【摘要】 为了研究禽流感病毒核蛋白(nucleoprotein,NP)在鸡体内所引起的免疫反应,将其NP基因从重组质粒pVAX-NP上酶切后亚克隆至含禽源启动子的真核表达载体pCAGGS中,经筛选鉴定后获得重组子pCAGGS-NP,将鉴定正确的阳性重组子体外转染真核细胞,利用间接免疫荧光法检测目的基因的瞬时表达;采用腿部肌肉注射途径将重组子免疫SPF鸡,利用ELISA检测免疫后血清抗NP蛋白抗体效价。结果表明,该重组子无论在体外还是体内均能正确表达禽流感NP蛋白,且所表达的蛋白均具有良好的免疫原性。更多还原

【Abstract】 In order to investigate the immune responses to the nucleoprotein(NP) of Avian influenza virus,NP gene cut from re-combinant plasmid pVAX-NP was subcloned into a eukaryotic expression vector pCAGGS with an avian-derived promoter.The re-combinant plasmid pCAGGS-NP was obtained after PCR identification and sequencing.Then the positive pCAGGS-NP was subject-ed to transfect eukaryocyte and to immunize SPF chickens.Indirect florescent assay(IFA) was engaged to detect the transient expres-sion of NP,and ELISA was used to detect the serum antibody of the immunized chickens which was resistant to NP protein.The re-sults showed that NP was successfully expressed both in vitro and in vivo and had good immunoreactivity.更多还原