摘要
[目的]实现PRRSV GP5与GP4蛋白在同一载体中表达各自编码的蛋白,发挥GP5蛋白在体液免疫中的优势和GP4蛋白在信号转导中的作用。[方法]利用RT-PCR扩增出GP5与GP4基因,克隆到pIRESneo载体的多克隆酶切位点及新霉素磷酸转移酶位点中,用XhoⅠ和NruⅠ双酶切含GP5和GP4基因的表达盒,将此表达盒克隆到犬2型腺病毒E3区缺失性载体pPolyⅡ-CAV-2-△E3中,以AsⅠc和PmeⅠ双酶切进行鉴定。[结果]将构建好的重组腺病毒免疫小鼠,可以诱导产生较强的体液免疫应答(ELISA抗体和中和抗体)。[结论]该重组腺病毒具有较好的免疫原性,可为研制有效的亚单位疫苗奠定基础。
[Objective] What the corresponding encoded protein of the PRRSV protein GP5 and GP4 could be expressed in same vector,respectively was achieved and the advantage role of protein GP5 in humoral immunity and protein GP4 in signal transduction was played.[Method] The gene GP5 and GP4,amplified with the RT-PCR,was cloned into the multiple-cloning enzyme restriction site of pIRESneo vector and the neomycin phosphotransferase loci;and the expression kit of gene GP5 and GP4 with double-digested enzyme of XhoⅠ and NruⅠwas cloned into the deficiency carrier pPolyⅡ-CAV-2-E3 in the E3 Zone of type 2 of canine adenovirus,which were identified with double-digested restriction enzymeof AscⅠ and PmeⅠ.[Results] A strong humoral immune response（ELISA antibodies and neutralizing antibodies） could be induced in the recombinant adenovirus immunized mice.[Conclusion] The recombinant adenovirus had good immunogenicity,by which the basis for developing an effective subunit vaccine would be laid.
出处
《安徽农业科学》
CAS
北大核心
2010年第10期5155-5157,5193,共4页
Journal of Anhui Agricultural Sciences
