CAJ | 학술논문

为原核表达猪繁殖与呼吸综合征病毒(PRRSV)的ORF3基因,本研究根据GenBank登录PRRSV美洲株ATCC VR2332的ORF3基因序列,利用Primet 6.0软件设计合成一对特异性引物.经RT-PCR扩增得到了大小为765 bp的片段.将扩增的ORF3基因截短为495 bp和750 bp片段分别克隆于原核表达栽体pGEX-KG中,在IPTG的诱导下进行Gp3重组蛋白的截短表达,经western blot检测证实表达的2种截短重组蛋白均具有良好的与抗体的反应活性,从而为进一步研制新型疫苗提供了实验依据.
위원핵표체저번식여호흡종합정병독(PRRSV)적ORF3기인,본연구근거GenBank등록PRRSV미주주ATCC VR2332적ORF3기인서렬,이용Primet 6.0연건설계합성일대특이성인물.경RT-PCR확증득도료대소위765 bp적편단.장확증적ORF3기인절단위495 bp화750 bp편단분별극륭우원핵표체재체pGEX-KG중,재IPTG적유도하진행Gp3중조단백적절단표체,경western blot검측증실표체적2충절단중조단백균구유량호적여항체적반응활성,종이위진일보연제신형역묘제공료실험의거.
For prokaryotic expression of Gp3 of protein of porcine reproductive and respiratory syndrome virus (PRRSV), the ORF3 gene from a PRRSV NC strain was amplified by RT-PCR using primers derived from PRRSV ORF3 gene encoding Gp3 protein of VR2332 strain. A 765 bp gene fragment was obtained and cloned into the prokaryotic expression vector pGEX-KG, The Gp3 recombinant protein was expressed in E.coli cells. Western blot test confirmed that the expressed recombinant protein could specifically react with the antiserum against PRRSV.