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。
Recombinant plasmid pET-PCP was constructed by ligating coat protein （CP） gene of Pennisetum mosaic virus（PenMV）into the expression vector pET22b（＋） and then transferred into E. coli BL21 （DE3） pLys. The results of SDS-PAGE and Western blotting analysis showed that about 36 kDa specific fusion protein was produced after induction by IPTG. The fusion protein was injected into rabbit to prepare antiserum after it was purified by Ni^2＋ -affinity chromatography. And the titer was determined to be 1 ： 1 024 by microprecipitation, or 1：8 192 by antigen coating plate-ELISA（ACP-ELISA）. The antiserum is useful for identification of PenMV in plant materials.
出处
《植物病理学报》
CAS
CSCD
北大核心
2005年第5期442-445,共4页
Acta Phytopathologica Sinica
基金
国家自然科学基金资助项目(30170608)
