CAJ | 학술논문

为获得猪瘟病毒（CSFV）E0重组蛋白，建立 CSFV 抗体快速检测方法。本研究通过扩增 CSFV C 株 E0基因，亚克隆至原核表达载体 pGEX-4T-2，构建重组原核表达质粒 pGEX-4T-2-E0，转化至宿主菌Rostta（DE3）后进行诱导表达。SDS-PAGE 检测表明与目的蛋白大小一致，其分子质量大小为46 ku。优化重组蛋白表达条件，最终获得诱导表达的最佳温度为37℃，最佳 IPTG 浓度为0．1 mmol/L，并且目的蛋白以包涵体形式存在。Western blot 表明，目的蛋白能与 CSFV 兔化高免血清反应，表明该融合蛋白具有良好的抗原性。本研究为 CSFV 抗体检测试剂盒的研究奠定了基础。
위획득저온병독（CSFV）E0중조단백，건립 CSFV 항체쾌속검측방법。본연구통과확증 CSFV C 주 E0기인，아극륭지원핵표체재체 pGEX-4T-2，구건중조원핵표체질립 pGEX-4T-2-E0，전화지숙주균Rostta（DE3）후진행유도표체。SDS-PAGE 검측표명여목적단백대소일치，기분자질량대소위46 ku。우화중조단백표체조건，최종획득유도표체적최가온도위37℃，최가 IPTG 농도위0．1 mmol/L，병차목적단백이포함체형식존재。Western blot 표명，목적단백능여 CSFV 토화고면혈청반응，표명해융합단백구유량호적항원성。본연구위 CSFV 항체검측시제합적연구전정료기출。
In order to gain the recombinant protein E0 of the classical swine fever virus(CSFV),to estab-lish CSFV antibody rapid detection method,the CSFV C strain E0 was amplified and sub-cloned into pro-karyotic expression plasmid pGEX-4T-2 vector.The recombinant plasmid named pGEX-4T-2-E0 was transformed into E.coli Rosetta (DE3)and induced by IPTG.The expression products were analyzed by SDS-PAGE,a specific expression band,in form of inclusion body with a molecular weight 46.0 ku was de-tected.The optimum temperature was 18℃,the IPTG was 1.0 mmol/L.Western-blot indicated that the expressed protein was recognized not only by the rabbit antibody to CSFV,but also with the specific mon-oclonal antibody of GST tag.The result suggests that the recombinant protein has good antigenicity and provides substantial base for the CSFV antibody detection kit research.