摘要
将AStV基因组组分3编码的NS3和NC蛋白基因以及组分4编码的NCP和NSvc4蛋白基因分别克隆于大肠杆菌表达载体pGEX3X在IPTG诱导下，表达出4种融合蛋白。这些表达产物用于免疫动物产生多克隆抗血清。以制备的多克隆抗血清为探针，Western印迹分析了宿主植物和介体昆虫体内NS3、NC、NCP和NSvc4种基因表达的产物。NC蛋白的抗体可在病株和病毒粒子制备物中以及NCP抗血清仅在病株中检测到相应大小的产物，其余的均与预期值相停。
The NS3 and NC protein genes encoded by RNA3 of RStV, the NCP and NSvc4protein genes encoded by RNA4 were subcloned into the E. coli expression vectorpGEX3X to express four groups of fusion protein under IPTG induction. These fusionproteins were used to immunize rabbits to raise antisera. The antisera against the Ecoli-expressed proteins were available for probing the presence of the viral geneproducts in both rice plant and insect hosts. The expected gene products can beprobed only in diseased rice plant with NCP antiserum and the cormsponding productsdetected in both plant and RStV panicle preparation with NC anhserum. The viralgene products probed by NS3 and NSvc4 anhsera were different from the expectedones in size.
基金
美国McKnight基金
上海市科委自然科学基金
关键词
融合蛋白
印迹分析
条叶枯病毒
水稻昆虫
Viral gene, Gene expression, Fusion protein, Blotting analysis
