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针对某些口蹄疫病毒非结构蛋白的血清学探针。

Serological probes for some foot-and-mouth disease virus nonstructural proteins.

作者信息

Tesar M, Berger H G, Marquardt O

机构信息

Federal Research Center for Virus Diseases of Animals, Tübingen, Federal Republic of Germany.

出版信息

Virus Genes. 1989 Sep;3(1):29-44. doi: 10.1007/BF00301985.

Abstract

Foot-and-mouth disease virus (FMDV) O1 Kaufbeuren-specific cDNA fragments were subcloned into the E. coli expression vector pRIT.2T. Fusion proteins thus produced in bacteria were purified by affinity chromatography and inoculated into rabbits. Three sera thus obtained were found to be monospecific for FMDV proteins 3A, 3C, and 3D, respectively. Two others were prevalently directed against protein 2C, but in addition, either to protein 2B or to protein 3A. Five out of six mature nonstructural virus proteins can therefore be separately investigated in FMDV-infected cells, either by indirect immunofluorescence or by radioimmunoprecipitation. Immunofluorescence shows all investigated proteins to be located exclusively in the cytoplasm. One of them, protein 2C, transiently forms aggregates at the periphery of cells. Radioimmunoprecipitation confirmed current knowledge on maturation of FMDV proteins. It was further used to characterize postinfectional sera with regard to FMDV-specific antibodies. Cattle and guinea pig were found to have responded differently to FMDV nonstructural antigens. Furthermore, antigenicity of yet to be described FMDV polypeptides was observed in the guinea pig.

摘要

口蹄疫病毒(FMDV)O1考夫博伊伦株特异性cDNA片段被亚克隆到大肠杆菌表达载体pRIT.2T中。在细菌中产生的融合蛋白通过亲和层析进行纯化,并接种到兔子体内。结果发现由此获得的三份血清分别对FMDV蛋白3A、3C和3D具有单特异性。另外两份血清主要针对蛋白2C,但此外,一份针对蛋白2B,另一份针对蛋白3A。因此,在FMDV感染的细胞中,可以通过间接免疫荧光或放射免疫沉淀法分别研究六种成熟的非结构病毒蛋白中的五种。免疫荧光显示所有研究的蛋白都仅位于细胞质中。其中一种蛋白2C在细胞周边短暂形成聚集体。放射免疫沉淀法证实了关于FMDV蛋白成熟的现有知识。它还被用于鉴定感染后血清中的FMDV特异性抗体。发现牛和豚鼠对FMDV非结构抗原的反应不同。此外,在豚鼠中观察到了尚未描述的FMDV多肽的抗原性。

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