Rodriguez María Jose, Sarraseca Javier, Fominaya Jesús, Cortés Elena, Sanz Antonio, Casal J Ignacio
INGENASA, Hermanos Garcia Noblejas 41, 28037 Madrid, Spain1.
J Gen Virol. 2001 May;82(Pt 5):995-999. doi: 10.1099/0022-1317-82-5-995.
Glycoprotein 5 (GP(5)) is the major glycoprotein of porcine reproductive and respiratory syndrome virus (PRRSV). Expression of GP(5) has been improved by removing the transmembrane regions. Vectors were constructed encoding complete GP(5) plus three mutants: GP(5) Ns (residues 28--201), GP(5)[30--67] (residues 30--67) and GP(5)[30--201] (residues 30--67/130--201). The three deletion mutants were expressed at levels 20--30 times higher than complete GP(5). GP(5)[30--201] was well recognized in ELISA or immunoblotting by a collection of pig sera. All the fragments were tested for the generation of MAbs, but only the polyhistidine-tagged fragment GP(5)[30--201]H elicited an antibody response sufficient to produce MABS: The two MAbs were positive for PRRSV in ELISA and immunoblotting, but negative for virus neutralization. MAb 4BE12 reacted with residues 130--170 and MAb 3AH9 recognized residues 170--201. This region was recognized strongly in immunoblotting by a collection of infected-pig sera. These results indicate diagnostic potential for this epitope.
糖蛋白5（GP(5)）是猪繁殖与呼吸综合征病毒（PRRSV）的主要糖蛋白。通过去除跨膜区域，GP(5)的表达得到了改善。构建了编码完整GP(5)以及三个突变体的载体：GP(5) Ns（第28至201位氨基酸残基）、GP(5)[30 - 67]（第30至67位氨基酸残基）和GP(5)[30 - 201]（第30至67位/第130至201位氨基酸残基）。这三个缺失突变体的表达水平比完整的GP(5)高20至30倍。在ELISA或免疫印迹中，一组猪血清能很好地识别GP(5)[30 - 201]。对所有片段进行了单克隆抗体（MAb）生成测试，但只有带有多聚组氨酸标签的片段GP(5)[30 - 201]H引发了足以产生单克隆抗体的抗体反应：这两种单克隆抗体在ELISA和免疫印迹中对PRRSV呈阳性，但在病毒中和试验中呈阴性。单克隆抗体4BE12与第130至170位氨基酸残基反应，单克隆抗体3AH9识别第170至201位氨基酸残基。在免疫印迹中，一组感染猪的血清强烈识别该区域。这些结果表明该表位具有诊断潜力。
