呼肠病毒BYD株吸附蛋白σ1的原核表达及其抗原性鉴定

Prokaryotic Expression and Antigenicity Analysis of the Attachment Protein σ1 of Reovirus BYD Strain

摘要:

将呼肠病毒BYD株S1片段编码的细胞吸附蛋白基因连接至pET-28a(+)，构建表达载体pET-28a(+)-S1，转化表达宿主菌E.coli BL21(DE3)，分析表达载体能否表达预期大小的重组σ1，并进一步鉴定重组σ1的抗原性。SDS-PAGE实验表明，经IPTG诱导后，表达载体能表达53kD左右蛋白质，与重组σ1的预期大小相符；免疫印迹分析表明重组σ1有较好的抗原性和特异性，可用于呼肠病毒诊断试剂的制备。

Abstract:

The cell attachment protein gene of the S1 segment of reovirus BYD was cloned into prokaryotic expression vector, pET-28a(+). The resulting pET-28a(+)-S1 was transformed into E.coli BL21(DE3) for analyzing whether the constructed vector could express inferred recombinant σ1 and characterizing the antigenicity of the recombinant σ1.SDS-PAGE showed the constructed vector expressed a protein around 53 kilodalton, which was consistent with the deduced molecular weight of the recombinant σ1.And immunoblot assays demonstrated that the recombinant σ1 had good antigenicity and specificity, and therefore it should have great potential in developing diagnosis reagent.