【摘要】 为得到可溶性表达的Cap蛋白并确定其抗原活性及存在形式,通过对IPTG浓度、诱导表达温度和时间进行优化,最终确定在OD600值为0.6～1.0时加入终浓度为1mmol/L的IPTG,37℃诱导4h为最佳的诱导表达条件。镍离子亲和层析对Cap蛋白进行纯化后,间接ELISA表明,重组Cap具有良好的抗原活性。非还原SDS-PAGE电泳分析表明,Cap重组蛋白除以26ku的单体分子存在外,还有一部分形成52ku的二聚体,为Cap病毒样颗粒的制备奠定基础。更多还原

【Abstract】 To obtain soluble expression of the recombinant Cap protein of PCV2 and analyze its immunogenicity,prokaryotic expression conditions were optimized.The optimal soluble expression of the recombinant Cap protein was obtained when E.coli BL21(DE3) cells harboring pET-28a-ORF2 were grown to log phase(OD600 = 0.6 to 1.0) and then induced by a final concentration of 1 mmol/L IPTG under 37 ℃ for 4 h.Following nickel ions chromatography,indirect ELISA showed that purified Cap protein possesses high immunogenicity.Non-reduced SDS-PAGE demonstrated that the recombinant Cap protein existed as both monomer of 26 ku and dimer of 52 ku.This study laid the foundation for preparing Cap-based virus like particles.更多还原