CAJ | 학술논문

为制备猪瘟病毒(CSFV)特异性诊断抗原,根据国内CSFV流行毒株的序列合成了其主要的保护性抗原Erns蛋白的基因序列,将合成的基因按设计的酶切位点与毕赤酵母表达载体pPIC9K连接,构建了重组表达质粒pPIC9K-Erns.用SalI将重组表达质粒线性化后,转化GS115酵母菌.经筛选、鉴定,成功获得了表达CSFV的Erns蛋白的阳性重组酵母菌.该阳性重组酵母菌经甲醇诱导,在培养液上清中可检测到目的蛋白.Western-blot结果显示,该蛋白能特异识别CSFV抗体,且具有很好的反应原性.
위제비저온병독(CSFV)특이성진단항원,근거국내CSFV류행독주적서렬합성료기주요적보호성항원Erns단백적기인서렬,장합성적기인안설계적매절위점여필적효모표체재체pPIC9K련접,구건료중조표체질립pPIC9K-Erns.용SalI장중조표체질립선성화후,전화GS115효모균.경사선、감정,성공획득료표체CSFV적Erns단백적양성중조효모균.해양성중조효모균경갑순유도,재배양액상청중가검측도목적단백.Western-blot결과현시,해단백능특이식별CSFV항체,차구유흔호적반응원성.
In order to produce the specifc diagnostic antigens of classical swine fever virus(CSFV),the gene sequence of CSFV's main protective antigen-Erns protein was synthesized according to the sequence of the prevalent CSFV Cstrain.Then the purified products were cloned into pPIC9K expression vector to construct recombinant expression plasmid(Ppic9k-Erns).After recombinant expression plasmid linearized by Sal I,the microzyme GS115 was transformed by Sal I.Through screening and identification,the positive recombinant micmzyme of Erns protein to express CSFV was obtained successfully.This positive recombinant microzyme induced by methanol,then,the target protein could be examined in the superuatant culture solution.The results of Western-blot showed that this target protein could specifically recognize the antigens of CSFV and it had a great reactionogenicity.