Martin A, Wychowski C, Benichou D, Crainic R, Girard M
Unité de Virologie Moleculaire UA CNRS 545, Institut Pasteur, Paris.
Ann Inst Pasteur Virol. 1988 Jan-Mar;139(1):79-88. doi: 10.1016/s0769-2617(88)80008-x.
A chimaeric poliovirus carrying a type-2-specific neutralization epitope on a type 1 capsid was created by site-directed mutagenesis of the Mahoney strain of poliovirus type 1. An EcoRV and a HindIII restriction sites were first constructed in the cDNA of poliovirus type 1 at nucleotide positions 2756 and 2786, respectively, i.e. on either side of the sequence encoding neutralization epitope C3 (VP1 amino acids 93-103), which is part of neutralization site NImI. The cDNA sequence framed by the two sites was next taken out and replaced by custom-made oligonucleotides encoding the equivalent region of VP1 from the Lansing strain of poliovirus type 2. The DNA from the plasmid carrying such a hybrid construct was transfected onto CV1 cells generating a chimaeric virus, v510. Neutralization of v510 with a panel of monoclonal antibodies showed that v510 has lost the poliovirus type 1 C3 epitope but acquired a new, poliovirus type-2-specific neutralization epitope. Preliminary results indicate that v510 also shows neurovirulence for mice, which is a specific trait of the Lansing strain of poliovirus type 2.
通过对1型脊髓灰质炎病毒Mahoney株进行定点诱变，构建了一种在1型衣壳上携带2型特异性中和表位的嵌合脊髓灰质炎病毒。首先在1型脊髓灰质炎病毒的cDNA中，分别于核苷酸位置2756和2786构建了EcoRV和HindIII限制性酶切位点，即位于编码中和表位C3（VP1氨基酸93 - 103）的序列两侧，该表位是中和位点NImI的一部分。接下来，取出由这两个位点界定的cDNA序列，并用编码2型脊髓灰质炎病毒Lansing株VP1等效区域的定制寡核苷酸进行替换。将携带这种杂交构建体的质粒DNA转染到CV1细胞上，产生了一种嵌合病毒v510。用一组单克隆抗体对v510进行中和试验表明，v510已失去1型脊髓灰质炎病毒的C3表位，但获得了一个新的、2型脊髓灰质炎病毒特异性中和表位。初步结果表明，v510对小鼠也表现出神经毒力，这是2型脊髓灰质炎病毒Lansing株的一个特异性特征。
