de Mattos C A, de de Mattos C C, Dangler C A, Osburn B I, Ianconescu M, Kaufmann R
Department of Veterinary Pathology, School of Veterinary Medicine, University of California, Davis.
Arch Virol. 1992;122(1-2):23-33. doi: 10.1007/BF01321115.
Partial cDNA clones representing 47%, 96%, and 98% of genome segments 7, 9, and 10, respectively, of a US bluetongue virus (BLU) 11 virulent strain were used to study, for the first time, the genetic relationships between Israeli BLU proto-serotypes and field isolates, and US BLU proto-serotypes. Their usefulness as group-specific identification probes was also determined. The viral nucleic acid was extracted from the infected cells and the purified dsRNA genome segments were fractionated by polyacrylamide gel electrophoresis, transferred to a nylon membrane and hybridized to the 32P labeled DNA probes. The three probes recognized all the samples tested. Genome segment 7, that code for the mayor inner capsid protein VP7, showed the most variation in the hybridization signal with the US proto-serotypes and all the Israeli samples studied. The genome segments 9 and 10 that code for the minor inner capsid protein VP6 and the nonstructural protein NS3, respectively, were highly conserved in all the samples tested despite their distant geographical regions of origin. The last two mentioned clones showed to be good group-specific probes for the identification of BLU samples from Israel and United States. The obtained cloned genetic probes were also tested against US epizootic haemorrhagic disease virus (EHDV) serotype 1 and 2 viral dsRNA, a distantly related orbivirus. None of them hybridized with the viral dsRNA of these two viruses.
分别代表美国蓝舌病病毒（BLU）11型强毒株基因组片段7、9和10的47%、96%和98%的部分cDNA克隆，首次被用于研究以色列BLU原型血清型与田间分离株以及美国BLU原型血清型之间的遗传关系。还确定了它们作为组特异性鉴定探针的效用。从感染细胞中提取病毒核酸，纯化的双链RNA基因组片段通过聚丙烯酰胺凝胶电泳进行分离，转移到尼龙膜上，并与32P标记的DNA探针杂交。这三种探针识别了所有测试样本。编码主要内衣壳蛋白VP7的基因组片段7，在与美国原型血清型和所有研究的以色列样本的杂交信号中显示出最大的变异性。分别编码次要内衣壳蛋白VP6和非结构蛋白NS3的基因组片段9和10，尽管其起源地地理位置遥远，但在所有测试样本中高度保守。最后提到的两个克隆显示是用于鉴定来自以色列和美国的BLU样本的良好组特异性探针。所获得的克隆遗传探针也针对美国 epizootic haemorrhagic disease virus（EHDV）血清型1和2病毒双链RNA进行了测试，这是一种远缘相关的环状病毒。它们均未与这两种病毒的病毒双链RNA杂交。
