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非洲马瘟病毒基因组的表征与克隆

Characterization and cloning of the African horsesickness virus genome.

作者信息

Bremer C W, Huismans H, Van Dijk A A

机构信息

Biochemistry Section, Veterinary Research Institute, Onderstepoort, South Africa.

出版信息

J Gen Virol. 1990 Apr;71 ( Pt 4):793-9. doi: 10.1099/0022-1317-71-4-793.

Abstract

The dsRNA profiles of all nine African horsesickness virus (AHSV) serotypes were compared by agarose gel electrophoresis and PAGE. The agarose profiles were identical, but a unique profile was obtained for each of the nine serotypes by PAGE. Nine of the 10 dsRNA genome segments of AHSV-3 were cloned and the clones were used in dot-spot and Northern blot hybridization experiments to determine intra- and inter-serogroup nucleic acid similarities. Segments 1, 3, 4, 5, 7 and 8 were highly conserved in the AHSV serogroup and no genetic relationship with any of the other orbiviruses was observed. Of these segments 3, 5 and 8 showed the largest degree of cross-hybridization to the cognate genes of all the serotypes. These clones did not cross-hybridize to other orbiviruses such as epizootic haemorrhagic disease virus, bluetongue virus or equine encephalosis virus and are therefore recommended for use as group-specific probes for the identification of the AHSV serogroup. Genome segments 6 and 10 showed an intermediate degree of conservation, whereas segment 2 is serotype-specific and therefore probably codes for the outer capsid protein VP2.

摘要

通过琼脂糖凝胶电泳和聚丙烯酰胺凝胶电泳(PAGE)对所有9种非洲马瘟病毒(AHSV)血清型的双链RNA(dsRNA)图谱进行了比较。琼脂糖凝胶电泳图谱相同,但通过PAGE获得了9种血清型各自独特的图谱。克隆了AHSV - 3的10个dsRNA基因组片段中的9个,并将这些克隆用于斑点杂交和Northern印迹杂交实验,以确定血清群内和血清群间核酸的相似性。片段1、3、4、5、7和8在AHSV血清群中高度保守,未观察到与其他环状病毒有任何遗传关系。在这些片段中,片段3、5和8与所有血清型的同源基因表现出最大程度的交叉杂交。这些克隆与其他环状病毒如流行性出血病病毒、蓝舌病病毒或马脑脊髓炎病毒不发生交叉杂交,因此推荐用作鉴定AHSV血清群的群特异性探针。基因组片段6和10表现出中等程度的保守性,而片段2是血清型特异性的,因此可能编码外衣壳蛋白VP2。

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