水泡性口炎病毒2种血清型核衣壳蛋白的原核表达及生物信息学分析
Prokaryotic Expression and Bioinformatics Analysis of the Nucleprotein Genes of Vesicular Stomatitis Virus
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摘要: 用RT-PCR方法从水泡性口炎病毒(vsv)扩增印第安纳(Indiana)和新泽西(New Jersey)血清型核蛋白N基因,分别克隆至pMD20-T载体进行序列鉴定及生物信息学分析.构建重组表达质粒VSV-IN-pBCX和VSV-NJ-pB-CX,并经SDS-PAGE和Western-blot鉴定表明2种血清型病毒核衣壳蛋白N基因在BL21(DE3)宿主菌中成功表达,重组蛋白相对分子质量约为82 000,并能够与各自对应血清型阳性血清反应.这表明2个重组蛋白具有良好的抗原性,可作为用于建立水泡性口炎血清学诊断方法的诊断抗原.Abstract: The nucleoprotein genes of vesicular stomatitis virus(VSV,Indiana type and New Jersey type) were amplified by reverse transcription polymerase chain reaction(RT-PCR),and then cloned into pMD20-T vector for sequencing and bioinformatics analysis.These genes were inserted into the prokaryotic expression plasmids pBCX.The recombinant plasmid VSV-IN-pBCX and VSV-NJ-pBCX were transformed into Escherichia.coli BL21(DE3) cells.The relative molecular mass of the recombinant protein was 82 000,which was detected by SDS-PAGE and reacted with VS positive serum in Western-blot.The recombinant nucleoproteins can be used as the specific diagnosis antigens for immunoassay such as ELISA.