作　　者： ; ; ; ; ; ;

机构地区： 沈阳农业大学园艺学院

出　　处： 《果树学报》 2013年第4期578-581,共4页

摘　　要： 【目的】探索以原核表达的草莓轻型黄边病毒(SMYEV)外壳蛋白(CP)为抗原制备抗血清的方法,从而建立利用ELISA检测SMYEV的技术体系。【方法】利用IPTG诱导SMYEV CP重组蛋白在大肠杆菌中表达,分离纯化重组蛋白并以其为抗原对新西兰兔进行免疫。血清效价达到要求后,分离纯化抗血清并利用DAC-ELISA对草莓植株进行SMYEV检测,同时利用RT-PCR对DAC-ELISA检测结果进行验证。【结果】宿有SMYEV CP基因的大肠杆菌经IPTG诱导后出现一条52.0 ku左右的特异性重组蛋白谱带,以纯化的重组蛋白为抗原制备出专一性较强的针对SMYEV的抗血清,该抗血清稀释800倍后仍能有效检测草莓植株中的SMYEV。利用DAC-ELISA和RT-PCR对26份草莓试材分别进行SMYEV检测,2种检测方法的检测结果基本一致。【结论】以原核表达的SMYEV CP为抗原可以制备出专一性较强、效价较高的SMYEV的抗血清。 [Objective] The objective of this study is to produce antiserum against strawberry mild yellow edge virus （SMYEV） with coat protein （CP） expressed in Escherichia coli cells as the antigen, thereby to develop the system of detection of SMYEV by ELISA. [Method] The recombinant coat protein of SMYEV was expressed in E. coli induced by IPTG, and the purified recombinant protein was used to raise anti- serum in the New Zealand rabbits. After the antiserum titer reached the standard, the antiserum was iso- lated and purified from the blood of rabbit and used to detect SMYEV in strawberry plants by DAC- ELISA. The test result of DAC-ELISA was verified by RT-PCR. [Result]A 52 kDa specific protein was induced by IPTG in E. coli. The purified recombinant protein raised the specific antiserum against SMYEV, which was still effective in detecting SMYEV by DAC-ELISA after diluted 800 folds. Twenty- six strawberry samples were detected for SMYEV by both DAC-ELISA and RT-PCR, and the test results of DAC-ELISA were basically consistent with that of RT-PCR. [Conclusion]The specific and high-titer antiserum against SMYEV was produced with CP expressed in prokaryotic cells as the antigen.

关 键 词： 草莓轻型黄边病毒 原核表达 抗血清

领　　域： [农业科学] [农业科学]