CAJ | 학술논문

为获得大量的猪瘟病毒重组蛋白抗原,本文构建了猪瘟兔化弱毒株E2蛋白ABCD抗原结构域原核重组表达质粒pET-32a（＋）-ABCD,对该重组表达质粒在大肠杆菌BL21（DE3）和Rosseta（DE3）中的表达产量进行了比较分析,并采用胶体金免疫层析和蛋白质免疫印迹鉴定重组蛋白的反应原性。结果表明重组质粒pET-32a（＋）-ABCD在大肠杆菌BL21（DE3）和Rosseta（DE3）中均可以获得可溶性表达,目的蛋白表达量分别占总蛋白量的比例为11.4%和19.7%,大肠杆菌Rosseta（DE3）作为表达宿主菌更高效,且重组蛋白具有反应原性。该研究为猪瘟病毒重组蛋白抗原的制备提供了参考。
위획득대량적저온병독중조단백항원,본문구건료저온토화약독주E2단백ABCD항원결구역원핵중조표체질립pET-32a（＋）-ABCD,대해중조표체질립재대장간균BL21（DE3）화Rosseta（DE3）중적표체산량진행료비교분석,병채용효체금면역층석화단백질면역인적감정중조단백적반응원성。결과표명중조질립pET-32a（＋）-ABCD재대장간균BL21（DE3）화Rosseta（DE3）중균가이획득가용성표체,목적단백표체량분별점총단백량적비례위11.4%화19.7%,대장간균Rosseta（DE3）작위표체숙주균경고효,차중조단백구유반응원성。해연구위저온병독중조단백항원적제비제공료삼고。
In order to obtain plenty of recombinant protein of classical swine fever virus,ABCD antigen region of E2 gene was amplified and used to construct pET-32a(+)-ABCD plasmid.The plasmid was then transformed into E.coli BL21(DE3) and Rosseta(DE3).The yields of expressed products in these two were compared as a reference to choose competent cells.The E2 protein was shown to be expressed in both E.coli BL21(DE3) and Rosseta(DE3) and accounted for 11.4% and 19.7% of the gross bacterial proteins as determined in SDS-PAGE.The recombinant E2 protein reacted with swine fever antiserum in gold immunochromatographic assay and western blotting.