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本试验成功构建了香蕉苞片花叶病毒(Banana bract mosaic virus,BBrMV)外壳蛋白(coat pro-tein,CP)基因的原核表达载体,并诱导表达了34 kDa的融合蛋白His.CP。对该原核表达蛋白的可溶性分析表明,该融合蛋白以包涵体形式存在。利用组氨酸标签纯化试剂盒对目的蛋白进行了纯化,获得了高纯度的融合蛋白。以纯化的蛋白为抗原免疫健康家兔,成功制备了抗BBrMV CP基因编码蛋白的兔抗血清。Western-blotting结果表明这种抗血清有很强的特异性。血清效价测定的效价在51 200倍以上,对植物材料的合适检测浓度为1:800～1:3 200。
The prokaryotic expression vector of coat pro-tein (CP) gene of banana bract mosaic virus (BBrMV) was successfully constructed and the fusion protein His.CP of 34 kDa was induced and expressed. Soluble analysis of the prokaryotic expressed protein showed that the fusion protein was in the form of inclusion bodies. The target protein was purified by histidine tag purification kit, and the fusion protein with high purity was obtained. Rabbit antiserum against BBrMV CP gene was successfully prepared by immunizing healthy rabbits with purified protein as antigen. Western-blotting results show that this antiserum has strong specificity. The titer of the serum titer is over 51 200 times, and the suitable test concentration for the plant material is 1: 800-1: 3 200.