中文摘要：

利用RT-PCR方法获得烟草蚀纹病毒（TEv）外壳蛋白（cP）基因，大小为789bp。经过EcoRI和NotI双酶切、定向克隆到pPIC9K，构建了真核表达载体pPIC9K．TEVCP。将重组质粒用sal I线性化后电转化导入毕赤酵母GS115中，经PCR鉴定为阳性的菌落，利用甲醇进行诱导表达，筛选出了高效表达菌株GS115-11和GS115-14，表达的蛋白大小约为37kD。表达产物经SDS．PAGE割胶纯化后，免疫家兔，制备了特异性抗血清，ELISA法检测效价为1：1500。Westernblot结果显示，制备的抗血清可以用来检测田间的发病植株。

英文摘要：

A TEV CP gene was obtained by means of RT-PCR amplification. The product was digested with restrictive endonu- clease Eco RI/Not I, and inserted into Pichia pastoris expression vector pPIC9K. Recombinant plasmid was linearized with Sal I and then transferred into GS115 strain by electroporation. Whether the TEV CP gene was integrated into alcohol oxidase promoter（AOX）locus on yeast chromosome was verified by amplification with special primers. SDS-PAGE result showed that the TEV CP gene was expressed highly in GS115-11 and GS115-14, and the molecular weight of expression product induced with 1% methanol was about 37 kD. The product was used as antigen to immunize the rabbit and the titer of antiserum was 1 ： 1500. Result of Western blot showed that the antiserum can be used to detect diseased tobaccos in the field.