猪瘟病毒E2蛋白抗原单表位基因的融合表达及其免疫活性的研究

利用基因搭桥技术，在Klenow DNA聚合酶作用下，分别合成猪瘟病毒E2蛋白的3个抗原表位基因，并与原核表达载体pGEX-3X进行连接、转化和筛选鉴定，构建了3个重组质粒pGEX-C、pGEX-D和pGEX-E。在IPTG的诱导下.实现了可溶性蛋白的融合表达(GST-C、GSTD、GST-E)，以GST亲和层析柱对融合蛋白进行纯化。应用ELISA和Western-blot检测证实：E抗原表位基因表达的融合蛋白GST-E具有免疫学活性，而C和D的抗原表位基因虽然都表达了融合蛋白GST-C和GSTD，但未检测到免疫学活性。免疫攻毒保护试验表明：融合蛋白GST-E具有一定的免疫保护功能，为多表位疫苗的研究奠定了基础。

Fusion Expression of Single-epitope Gene of CSFV Envelope Glycoprotein E2 and its Immunity Efficiency

The three epitope genes of CSFV envelope glycoprotein E2 were synthesized using recursive polymerization method, and three recombinant plasmids pGEX-C, pGEX-D and pGEX-E were constructed respectively. The three fusion proteins GST-C, GST-D and GST-E were expressed in E.coli BL21 and purified by glutathione resin volume. Immunological activity of the three fusion proteins were analyzed by ELISA and Western blot. The results showed that all the three fusion proteins could interacted with pig antisera against CSFV, however, only GST-E can strongly interacted with rabbit antisera against E2. The immunized rabbits were challenged with 100 MID_(50) hog cholera lapinized virus(HCLV), the result showed that the body temperature of rabbits of the control group and the vector protein-immunized group increased 24 hours postchallenge and didn't recovered to normal until 54 hours after challenge. However, the single epitope-immunized rabbits showed a mild increase of body temperature. This study will greatly promote the research of epitope vaccine of CSFV.