摘要苦苣菜黄网病毒(Sonchus yellow net virus，SYNV)是一种细胞核内复制的负链RNA病毒，它侵染菊科植物，在莴苣等农作物上造成病毒病害。本实验以纯化的重组蛋白为抗原免疫BALB/c小鼠，获得2株高特异性的单克隆抗体3C7和4D1。单抗亚类检测表明3C7细胞株单抗的重链和轻链类型分别是IgG2a和Lamda型，而4D1细胞株单抗的重链和轻链类型则分别为IgG2b和Lamda型。当包被原为3ug/ml时，3C7和4D1细胞株单抗腹水的效价分别为1:64000和1:128000。P蛋白单抗的研制为SYNV病毒的检测，以及P蛋白与寄主和病毒的其他蛋白的互作研究打下基础。46796

 Sonchus yellow net virus (SYNV) is a negative-strand RNA virus that replicates in the nucleus. SYNV infects composite plants and leads to viral diseases in some crops such as lettuce. The purified recombinant P was used to immunize BALB/c mice for producing MAbs. Two monoclonal antibodies named 3C7 and 4D1 were obtained. The results of monoclonal antibody subtypes detecting revealed the types of heavy chain and light chain of MAb from cell lines 3C7  were IgG2a and Lamda, while those of MAb from cell lines 4D1 were IgG2b and Lamda. When coating antigen concentration of 3ug/ml, the ELISA titres of ascitic fluids of MAbs from cell lines 3C7 and 4D1 were 1:64000 and 1:128000. Monoclonal antibodies preparation of sonchus yellow net virus P protein provided a foundation to study on the interactions between P proteins and host proteins, as well as the other proteins of SYNV. 

毕业论文关键词：苦苣菜黄网病毒；P蛋白；单克隆抗体。

Keywords: Sonchus yellow net virus; P protein; Monoclonal antibodies.

目录

第一章 病毒结构 4

1.1 植物弹状病毒 4

1.1.1 病毒粒子的形态和组成 4

1.1.2 基因组结构和组成 5

1.2 苦苣菜黄网病毒 5

1.2.1 基因组结构 5

1.2.2 结构蛋白功能 5

第二章 P蛋白单克隆抗体的制备 6

2.1 引言 6

2.2 材料 7

2.2.1 实验动物 7

2.2.2  细胞 7

2.2.3  化学试剂盒仪器 7

2.3 方法 7

2.3.1蛋白含量标准曲线的绘制 7

2.3.2 P蛋白单克隆抗体的制备 8

2.3.2.1乳化免疫原 8

2.3.2.2 免疫小鼠 8

2.3.2.3 抗血清效价测定 8

2.3.2.4 复苏骨髓瘤细胞 9

2.3.2.5  细胞融合 9

2.3.2.6杂交瘤细胞的筛选与克隆 9

2.3.2.7冻存与复苏细胞 10

2.3.2.8 单抗制备 10

2.3.2.9 纯化单抗 11

2.3.2.10  单抗浓度和效价测定 11

2.4 结果与分析 11

2.5 讨论 15

参考文献： 16

致谢 18

第一章 病毒结构

1.1 植物弹状病毒