【摘要】 将白草花叶病毒(Penn isetum mosaic virus,PenMV)的外壳蛋白(CP)基因连接到表达载体pET22b(+)上,获得重组子pET-PCP。SDS-PAGE和W estern b lotting分析表明,经IPTG诱导后,含pET-PCP的大肠杆菌BL21(DE3)pLys产生分子量为36 kDa的融合蛋白。以N i2+亲和层析柱纯化该蛋白,并以其为抗原免疫德国大白兔制备了特异性较强的抗血清。微量免疫沉淀法测其效价为1∶1 024,酶联法(enzym e-linked immunosorbant assay,ELISA)测定的效价为1∶8 192。该血清可替代常规病毒粒子所制备的抗血清,用于检测PenMV。更多还原

【Abstract】 Recombinant plasm id pET-PCP was constructed by ligating coat protein(CP) gene of Pennisetum mo-saic virus(PenMV) into the expression vector pET22b(+) and then transferred intoE.coliBL21(DE3) pLys.The results of SDS-PAGE andW estern blotting analysis showed that about 36 kDa specific fusion protein was pro-duced after induction by IPTG.The fusion protein was injected into rabbit to prepare antiserum after itwas puri-fied by N i2+-affinity chromatography.And the titerwas determ ined to be 1∶1 024 by m icro-precipitation,or 1∶ 8 192 by antigen coating plate-ELISA(ACP-ELISA).The antiserum is useful for identification of PenMV in plant materials.更多还原