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Polyclonal antibodies to a recombinant coat protein of potato virus A

机译:马铃薯A病毒重组外壳蛋白多克隆抗体

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Specific mouse antibodies against a recombinant coat protein (CP) of Potato virus A (PVA) were produced. The PVA CP gene was cloned in an expression vector pMPM4Omega. After expression in Escherichia coli the presence of the expressed CP was proved by Western blot analysis using polyclonal and monoclonal antibodies (MAbs). The expressed CP was purified by centrifugation in CsCl density gradient or on a sucrose cushion. The production of virus-like particles (VLPs) was proved by electron microscopy. The purified CP was used for preparation of a mouse antiserum which had a titer of 1: 1024 in ELISA and reacted specifically in Western blot analysis and indirect plate-trapped antigen ELISA (PTA-ELISA).
机译:产生了针对马铃薯病毒A(PVA)重组外壳蛋白(CP)的特异性小鼠抗体。将PVA CP基因克隆到表达载体pMPM40Omega中。在大肠杆菌中表达后,使用多克隆抗体和单克隆抗体(MAb)通过蛋白质印迹分析证实了表达的CP的存在。通过在CsCl密度梯度中或在蔗糖垫上离心来纯化表达的CP。通过电子显微镜证实了病毒样颗粒(VLP)的产生。纯化的CP用于制备小鼠抗血清,该抗血清在ELISA中的效价为1:1024,并在Western blot分析和间接板捕获的抗原ELISA(PTA-ELISA)中发生特异性反应。

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