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通过交叉杂交和交叉免疫沉淀法对澳大利亚蓝舌病毒分离株(CSIRO 19)与其他蓝舌病毒血清型进行比较。

A comparison of an australian bluetongue virus isolate (CSIRO 19) with other bluetongue virus serotypes by cross-hybridization and cross-immune precipitation.

作者信息

Huismans H, Bremer C W

出版信息

Onderstepoort J Vet Res. 1981 Jun;48(2):59-67.

PMID:6273774
Abstract

No major differences in size were observed when both the double-stranded RNA and the polypeptides of the Australian bluetongue virus (BTV) isolate CSIRO 19 (BTV-20) were compared with those of other BTV serotypes such as BTV-10 and BTV-4. Minor capsid polypeptide P6 of both BTV-20 and BTV-4, which electrophoreses as a single band on continuous phosphate buffered gels, in separated into 2 distinct bands on discontinuous glycine-buffered gels. This was not the case with BTV-10. Cross-immune precipitation of BTV-20 with BTV-10, BTV-17, BTV-4 and BTV-3 indicated strong immunological cross-reaction of the group-specific antigen P7 of the different serotypes. There was also some cross-immune precipitation of the serotype-specific polypeptide P2 of BTV-20 and BTV-4. This result is in agreement with the observed cross neutralization of these 2 viruses. The main distinction between BTV-20 and the other BTV serotypes was observed in cross-hybridization experiments. The homology between the nucleic acid of BTV-20 and other BTV serotypes was less than 30%, whereas homology normally found between BTV serotypes is at least 70%. The hybridization products of the different BTV serotypes were analysed by electrophoresis and fluorography. Two main hybrid segments were observed in all heterologous hybridizations with BTV-20 as a compared with 7 hybrid segments in hybridizations between BTV-4 and BTV-10. In order to determine from which genome segment of BTV-20 these 2 hybrid segments were derived, the hybridizations were carried out with individually purified double-stranded RNA segments. These results indicate that the 2 segments of BTV-20 that show the largest homology to corresponding segments of a heterologous BTV serotype are No. 7 and 10.

摘要

当将澳大利亚蓝舌病毒(BTV)分离株CSIRO 19(BTV - 20)的双链RNA和多肽与其他BTV血清型(如BTV - 10和BTV - 4)进行比较时,未观察到大小上的主要差异。BTV - 20和BTV - 4的次要衣壳多肽P6在连续磷酸盐缓冲凝胶上电泳时呈现为一条带,而在不连续甘氨酸缓冲凝胶上则分离为两条不同的带。BTV - 10并非如此。用BTV - 10、BTV - 17、BTV - 4和BTV - 3对BTV - 20进行交叉免疫沉淀表明,不同血清型的群特异性抗原P7存在强烈的免疫交叉反应。BTV - 20和BTV - 4的血清型特异性多肽P2也存在一些交叉免疫沉淀。这一结果与观察到的这两种病毒的交叉中和现象一致。在交叉杂交实验中观察到了BTV - 20与其他BTV血清型之间的主要区别。BTV - 20与其他BTV血清型核酸之间的同源性小于30%,而BTV血清型之间通常的同源性至少为70%。通过电泳和荧光自显影分析了不同BTV血清型的杂交产物。与BTV - 4和BTV - 10之间杂交产生7条杂交片段相比,以BTV - 20作为对照的所有异源杂交中均观察到两条主要的杂交片段。为了确定BTV - 20的这两条杂交片段源自哪个基因组片段,用单独纯化的双链RNA片段进行杂交。这些结果表明,BTV - 20中与异源BTV血清型相应片段显示出最大同源性的两个片段是7号和10号。

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