薤花叶病毒外壳蛋白抗血清制备及其与相关病毒血清学关系

Preparation of antiserum to the coat protein of Scallion mosaic virus and its serological relationships to other potyviruses

为研究薤花叶病毒(Scallionmosaicvirus,ScaMV)与其它一些马铃薯Y病毒属成员之间的血清学关系,先构建原核表达载体pSB-ScaMV-CP,然后确定该质粒能在大肠杆菌BL21plysS过量表达目的蛋白,最后用两次制备SDS-PAGE电泳分离纯化的CP免疫小鼠,获得的抗血清效价为1∶1024。Westernblot分析表明,ScaMVCP抗血清能与病毒自身CP有较强的特异性反应,与芜菁花叶病毒(TuMV)、水仙黄条病毒(NYSV)、小西葫芦黄花叶病毒(ZYMV)和芋花叶病毒(DsMV)CP有较弱的反应,与其它15种马铃薯Y病毒属病毒CP无反应。

The complete coat protein gene of Scallion mosaic virus from wild scallion in Hangzhou City, Zhejiang Province was amplified by PCR with specific primers and then inserted into pSBET. The recombinant plasmid of pSB-ScaMV-CP was transformed to BI221 plys S, in which the aimed protein was overexpressed after induced by IPTG and then purified by SDS-PAGE. The titre of antiserum obtained in the mice was 1：1 024 assayed by indirect ELISA. Based on Western blot analysis, the polycloned antiserum obtained reacted strongly and specifically to its homologous protein. It also reacted less strongly to the prokaryotically expressed CPs of Turnip mosaic virus, Narcissus yellow stripe virus, Zucchini yellow mosaic virus, and Dasheen mosaic virus, but not to other 15 potyviruses.