猪圆环病毒4型Cap蛋白多克隆抗体的制备OA北大核心

Using the synthesized full-length genomic DNA of PCV4 as a template,the genes PCV4 Cap and dCap were amplified and cloned into the prokaryotic expression vector pET-28a to obtain recombinant plasmids pET-28a-PCV4-Cap and pET-28a-PCV4-dCap.After digestion and sequencing,the recombinant plasmids were transformed into E.coli BL21 competent cells.After induction by IPTG,the fusion protein His-PCV4-dCap of nearly 28 kDa was successfully expressed and purified by Ni-NTA resin affinity chro-matography.BALB/c mice were immunized three times by subcutaneous multi-point injection to prepare spe-cific polyclonal antibody against PCV4 Cap protein.Western-blotting and indirect immunofluorescence(IFA)results showed that the polyclonal antibody exhibited high reactivity with prokaryotic and eu-karyotic expression of PCV4 Cap.The polyclonal antibody against PCV4 Cap protein prepared in this study lays a foundation for establishing a rapid epidemiological investigation of PCV4 infection,and studying the function of PCV4 Cap and revealing the mechanism of PCV4 pathogenicity.