Bruneau P, van der Werf S, Wychowski C, Siffert O, Girard M
Dev Biol Stand. 1984;57:177-85.
Poliovirus cDNA sequences encoding type 1 capsid polypeptide VP1 were fused in phase into the beta-lactamase sequence of pBR322. The resulting recombinant plasmid pSW119 expressed in E. coli a VP1-beta-lactamase fusion protein which reacted with antibodies raised against poliovirus capsid polypeptide VP1 and with a monoclonal poliovirus type 1 neutralizing antibody C3. Deletions of various length were introduced into the VP1 sequence. The truncated proteins expressed by the deleted plasmids did not react further with C3 when the region of VP1 amino acids 91-104 was deleted. A synthetic peptide corresponding to this region of VP1 (amino acids 93-104) was chemically synthesized. It was found to inhibit the seroneutralization of poliovirus by C3 antibodies and, once coupled to keyhole limpet hemocyanin, it was specifically immunoprecipitated with C3. Therefore, the C3 epitope recognized by the poliovirus neutralizing monoclonal antibody is located within the region of amino acids 93-104 of capsid polypeptide VP1.
编码1型衣壳多肽VP1的脊髓灰质炎病毒cDNA序列与pBR322的β-内酰胺酶序列同框融合。所得重组质粒pSW119在大肠杆菌中表达一种VP1-β-内酰胺酶融合蛋白，该融合蛋白能与针对脊髓灰质炎病毒衣壳多肽VP1产生的抗体以及一种脊髓灰质炎病毒1型中和单克隆抗体C3发生反应。对VP1序列进行了不同长度的缺失。当VP1氨基酸91 - 104区域缺失时，缺失质粒表达的截短蛋白不再与C3发生反应。合成了一段对应于VP1该区域（氨基酸93 - 104）的合成肽。发现它能抑制C3抗体对脊髓灰质炎病毒的血清中和作用，并且一旦与钥孔血蓝蛋白偶联，就能被C3特异性免疫沉淀。因此，脊髓灰质炎病毒中和单克隆抗体识别的C3表位位于衣壳多肽VP1的氨基酸93 - 104区域内。
