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首页> 外文期刊>Clinical and diagnostic laboratory immunology >Importance of M-Protein C Terminus as Substrate Antigen for Serodetection of Equine Arteritis Virus Infection
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Importance of M-Protein C Terminus as Substrate Antigen for Serodetection of Equine Arteritis Virus Infection

机译：M蛋白C总站作为底物抗原对马动脉炎病毒感染的血清学检测的重要性

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Equine arteritis virus (EAV), an enveloped positive-stranded RNA virus, is the prototype of the arterivirus group. In a previous paper (A. Kheyar, S. Martin, G. St.-Laurent, P. J. Timoney, W. H. McCollum, and D. Archambault, Clin. Diagn. Lab. Immunol. 4:648-652, 1997), we have shown that the unglycosylated membrane (M) protein, which is composed of 162 amino acids (aa), is a major target of equine antibody to EAV. In order to determine the antigenic regions of the M protein, the cDNA encoding the M protein of EAV was inserted into the procaryotic expression vector pGEX-4T-1 to produce recombinant glutathione S-transferase-M fusion protein. Various deletion mutant clones, which covered the entire sequence of the M protein, were then generated by inverse PCR and expressed in Escherichia coli to examine, by a Western blot assay, the antigenic reactivity of the clone-derived truncated M proteins with sera from horses either experimentally or naturally infected with EAV. Deletion of the hydrophobic N-terminal 87 aa did not abolish immune reactivity of the protein with serum antibodies to EAV, thereby demonstrating the antigenicity of the C-terminal region (aa 88 to 162) of the M protein. Further truncations of the M-protein C-terminal domain defined particular linear epitope-containing amino acid sequence regions. However, only the M-protein C-terminal region was readily recognized by all EAV-specific horse antisera tested in this study. Based on these findings, only the M-protein C-terminal polypeptide composed of aa 88 to 162 is necessary to identify horse serum antibodies specific to the EAV M protein. Thus, this polypeptide might be useful for serodetection of EAV-infected animals.
机译：马动脉炎病毒（EAV）是一种包膜的正链RNA病毒，是该动脉病毒组的原型。在以前的论文中（A. Kheyar，S。Martin，G。St.-Laurent，PJ Timoney，WH McCollum和D. Archambault，Clin。Diagn。Lab。Immunol。4：648-652，1997），我们有结果表明，由162个氨基酸（aa）组成的未糖基化膜（M）蛋白是针对EAV的马抗体的主要靶标。为了确定M蛋白的抗原区域，将编码EAV M蛋白的cDNA插入原核表达载体pGEX-4T-1中，以生产重组谷胱甘肽 S -转移酶-M融合蛋白。 。然后，通过反向PCR产生覆盖M蛋白整个序列的各种缺失突变体克隆，并在大肠杆菌中表达，以通过Western blot测定来检查该克隆衍生的抗原反应性。通过实验或自然感染EAV的马血清截断了M蛋白。疏水性N末端87aa的缺失并未消除该蛋白质与针对EAV的血清抗体的免疫反应性，从而证明了M蛋白质的C末端区域（aa 88至162）的抗原性。 M-蛋白C-末端结构域的进一步截短定义了特定的含线性表位的氨基酸序列区域。但是，在这项研究中测试的所有EAV特异性马抗血清仅能轻易识别M蛋白C末端区域。基于这些发现，仅需要由aa 88至162组成的M蛋白C末端多肽来鉴定对EAV M蛋白具有特异性的马血清抗体。因此，该多肽可用于血清检测EAV感染的动物。

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