Lettuce necrotic yellows virus (LNYV) is a plant rhabdovirus which has a type-1 transmembrane glycoprotein. Here, we describe the generation of murine anti-sera to the glycoprotein. Rational design, expression, and purification of recombinant glycoprotein, termed rLGe, was undertaken using SignalP4.1 and camSol servers to predict signal peptide cleavage and protein solubility. In order to successfully obtain expression in mammalian cells, LNYV glycoprotein native signal peptide was substituted with that of Rabies virus glycoprotein. In addition, rather than expression of the entire molecule, rLGe consisted of the LNYV glycoprotein ectodomain fused to two affinity tags to minimize the risk of protein aggregation, while facilitating detection and purification. rLGe was transiently expressed in mammalian cell culture, purified using affinity column chromatography, and used to immunize mice. Harvested anti-sera were immunoreactive and specific to the naturally occurring glycoprotein as confirmed by western blotting of plant leaf tissue infected with LNYV.

生菜坏死黄病毒（LNYV）是一种植物弹状病毒，具有1型跨膜糖蛋白。在这里，我们描述了鼠抗糖蛋白血清的产生。使用SignalP4.1和camSol服务器进行了重组糖蛋白（称为rLGe）的合理设计，表达和纯化，以预测信号肽的裂解和蛋白溶解度。为了成功获得在哺乳动物细胞中的表达，用狂犬病病毒糖蛋白取代了LNYV糖蛋白天然信号肽。另外，rLGe并非由整个分子表达，而是由LNYV糖蛋白胞外域与两个亲和标签融合而成，以最大程度地降低蛋白质聚集的风险，同时促进检测和纯化。rLGe在哺乳动物细胞培养物中瞬时表达，使用亲和柱色谱法纯化，并用于免疫小鼠。收获的抗血清具有免疫反应性，并且对天然存在的糖蛋白具有特异性，这一点已通过感染LNYV的植物叶片组织的蛋白质印迹进行了证实。