CAJ | 학술논문

为获得猪瘟病毒(classical swine fever virus, CSFV) NS2-3抗原集中区蛋白,并建立CSFV抗体快速检测方法.本研究以CSFV全长基因组质粒为模板,PCR扩增NS2-3抗原表位集中区,利用扩增片段和克隆载体,构建重组表达质粒,命名为pET32a-NS2-3-1.重组表达质粒转化Rosetta (DE3)细胞,利用IPTG诱导表达, SDS-PAGE电泳和Western-blot鉴定重组表达产物.结果表明,重组质粒pET32a-NS2-3-1在28℃诱导5 h得到高效表达,重组蛋白能够与兔抗CSFV阳性血清发生反应.获得CSFV NS2-3抗原集中区蛋白,并且获得的重组蛋白具有抗原性,能够作为CSFV抗体检测的抗原.
위획득저온병독(classical swine fever virus, CSFV) NS2-3항원집중구단백,병건립CSFV항체쾌속검측방법.본연구이CSFV전장기인조질립위모판,PCR확증NS2-3항원표위집중구,이용확증편단화극륭재체,구건중조표체질립,명명위pET32a-NS2-3-1.중조표체질립전화Rosetta (DE3)세포,이용IPTG유도표체, SDS-PAGE전영화Western-blot감정중조표체산물.결과표명,중조질립pET32a-NS2-3-1재28℃유도5 h득도고효표체,중조단백능구여토항CSFV양성혈청발생반응.획득CSFV NS2-3항원집중구단백,병차획득적중조단백구유항원성,능구작위CSFV항체검측적항원.
To identify the main antigen domain of the classical swine fever virus (CSFV) NS2-3, a method of detecting antibody of classical swine fever virus was established. CSFV genome plasmid as PCR template, the main antigenic domain of CSFV NS2-3 was amplified by PCR, amd PCR products was cloned into prokaryotic expression plasmid pET-32a(+) vector to obtain the recombinant expression plasmid, named as pET32a-NS2-3-1. The recombinant expression plasmid was transformed into E. coli Rosetta (DE3) and inducible expression by IPTG. The expression products were analyzed by SDS-PAGE and identified by Western-blot. The recombinant plasmid pET32a-NS2-3-1 was highly expressed, induced 5 hours could get a large number of recombinant protein at 28℃, and the recombinant protein reacted strongly with the C-Positive serum of CSFV. The main antigenic domain protein in NS2-3 of CSFV was obtained, which has good antigenicity for detecting antibodies against CSFV.