CAJ | 학술논문

为获得蓝舌病病毒(BTV)VP 7基因原核表达蛋白并检测其生物学活性,根据Genbank登录的BTVVP 7基因,设计1对引物,运用RT-PCR扩增技术获得BTVVP 7目的片段,然后定向克隆到 pPROEXHTb原核表达载体上,再转化重组质粒 pPROEXHTb-VP7到BL21感受态细胞,IPTG 诱导,表达产物纯化,进行 SDS-PAGE 鉴定及 Western-blot反应原性分析。结果表明：表达的 VP7目的蛋白大小为35 ku,Western-blot 分析表明纯化后的蛋白可以与BTV单克隆抗体发生特异性反应。
위획득람설병병독(BTV)VP 7기인원핵표체단백병검측기생물학활성,근거Genbank등록적BTVVP 7기인,설계1대인물,운용RT-PCR확증기술획득BTVVP 7목적편단,연후정향극륭도 pPROEXHTb원핵표체재체상,재전화중조질립 pPROEXHTb-VP7도BL21감수태세포,IPTG 유도,표체산물순화,진행 SDS-PAGE 감정급 Western-blot반응원성분석。결과표명：표체적 VP7목적단백대소위35 ku,Western-blot 분석표명순화후적단백가이여BTV단극륭항체발생특이성반응。
A pair of primers for theVP7 gene of bluetongue virus (BTV)was designed based on the se-quences from Genbank.RT-PCR was used to amplify the VP7 fragment of BTV,and then the fragment was subcloned to a prokaryotic expression vector pPROEXHTb to construct the targeting vector pPRO-EXHTb-VP7.Targeting vector was transformed to BL21 competent bacteria,and IPTG was added to in-duce the expression of VP7 protein.The protein was identified after purification SDS-PAGE and Western-blotting.The results showed that the molecular weight of VP7 was 35 ku,and the purified protein could re-act specifically to VP7 monoclonal antibodies.