Adam G, Heegard P, Bøg-Hansen T C, Mundry K W
Institute for Biology, University of Stuttgart, F.R.G.
J Virol Methods. 1987 Sep;17(3-4):263-75. doi: 10.1016/0166-0934(87)90136-4.
Thirteen different, biotinylated plant lectins were tested for their ability to recognize specifically the glycoproteins of the two different plant rhabdoviruses potato yellow dwarf virus and eggplant mottled dwarf virus. All viruses were propagated on the same plant host species, Nicotiana rustica L. The lectin-binding to the viral proteins was tested after electrophoretic separation and transfer to nitrocellulose membranes. Besides purified virus also partially pure virus preparations were used for the tests, in order to determine the specificity. The lectins had been selected for specificities to either one of the following monosaccharides: mannose, glucose, galactose, N-acetyl-D-galactosamine, N-acetyl-D-glucosamine and fucose. In the test panel of thirteen lectins, seven were found to react with the viral glycoproteins. Among these, four (LCA, VFA, PSA, Con A) belonged to the mannosyl- or glycosyl-specific group. However, these four lectins reacted also with other host proteins when partially pure virus preparations were used as samples. The other three lectins (GSA2b, STA, WGA) were specific for N-acetyl-D-glucosamine and detected almost exclusively the viral glycoproteins. Two of these lectins, STA and WGA, were extremely suitable for virus-specific assays, since they did not react with glycoproteins in healthy controls that were identical or comparable in their electrophoretic mobility with the rhabdovirus glycoproteins. No binding to viral glycoproteins was observed with galactose-, N-acetyl-galactosamine- and fucose-specific lectins. The assay for rhabdovirus glycoproteins in plants with the lectins was approximately 8-16 times less sensitive than with virus-specific antibodies.
测试了13种不同的生物素化植物凝集素识别两种不同植物弹状病毒——马铃薯黄矮病毒和茄子斑驳矮化病毒糖蛋白的能力。所有病毒均在同一植物宿主物种黄花烟草上繁殖。在电泳分离并转移至硝酸纤维素膜后，检测凝集素与病毒蛋白的结合情况。除了纯化病毒外，还使用部分纯化的病毒制剂进行测试，以确定其特异性。这些凝集素已根据对以下单糖之一的特异性进行选择：甘露糖、葡萄糖、半乳糖、N-乙酰-D-半乳糖胺、N-乙酰-D-葡萄糖胺和岩藻糖。在13种凝集素的测试组中，发现7种与病毒糖蛋白发生反应。其中，4种（扁豆凝集素、荆豆凝集素、花生凝集素、刀豆球蛋白A）属于甘露糖基或糖基特异性组。然而，当使用部分纯化的病毒制剂作为样品时，这4种凝集素也与其他宿主蛋白发生反应。另外3种凝集素（麦胚凝集素2b、大豆凝集素、小麦胚凝集素）对N-乙酰-D-葡萄糖胺具有特异性，几乎只检测到病毒糖蛋白。其中两种凝集素，大豆凝集素和小麦胚凝集素，非常适合用于病毒特异性检测，因为它们在健康对照中不与电泳迁移率与弹状病毒糖蛋白相同或相当的糖蛋白发生反应。未观察到半乳糖特异性、N-乙酰半乳糖胺特异性和岩藻糖特异性凝集素与病毒糖蛋白的结合。用凝集素检测植物中弹状病毒糖蛋白的方法比用病毒特异性抗体检测的灵敏度低约8-16倍。
