cDNA-cloning and expression of VP1-specific sequences of foot-and-mouth disease virus types A5 and O1.

L Meyer, H W Heinrich, M Lenk, S Kleinwächter, H Horstmann, S Bochnig, P Eichler, P Kruschke
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Abstract

The RNA genome of foot- and mouth disease virus strains A5 Westerwald and O1 Lausanne has been reverse-transcribed and cloned in lambdaphages or plasmids. Identification of cDNA-clones containing VP1-specific sequences was achieved by hybridization, restriction mapping, and sequence analysis. VP1-coding cDNA-fragments were subcloned into the expression vector pEX which led to synthesis of fusion proteins with beta-galactosidase. These fusion proteins reacted with anti-VP1 antibodies on a Western blot, but were not capable of inducing neutralizing antibodies to mice. This seemed to suggest a tertiary structure of the VP1-epitopes unlike those of native VP1. Other attempts are discussed to construct VP1-fusion proteins folding similarly to the native viral protein structure.

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口蹄疫病毒A5型和O1型vp1特异性序列的cdna克隆与表达
对口蹄疫病毒A5 Westerwald株和O1 Lausanne株的RNA基因组进行了逆转录,并在噬菌体或质粒中进行了克隆。通过杂交、限制性内切作图和序列分析,鉴定了含有vp1特异性序列的cdna克隆。将编码vp1的cdna片段亚克隆到表达载体pEX中,与β -半乳糖苷酶合成融合蛋白。这些融合蛋白在Western blot上与抗vp1抗体反应,但不能诱导小鼠产生中和抗体。这似乎表明VP1表位的三级结构与天然VP1表位不同。其他尝试讨论构建vp1融合蛋白折叠类似于天然病毒蛋白结构。
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