ISOLATION AND CHARACTERIZATION OF ORBIVIRUS GENOTYPIC VARIANTS
ISOLATION AND CHARACTERIZATION OF ORBIVIRUS GENOTYPIC VARIANTS
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DOI：
10.1016/0168-1702(87)90067-0
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发表时间：
1987-01-01
期刊：
影响因子：
5
通讯作者：
GOULD, AR
中科院分区：
文献类型：
--
作者：
EATON, BT;GOULD, AR
已经分离出含有RNA缺失或串联RNA的环状病毒变体。茨木病毒（鹿血清群的动物流行性出血性疾病的一个成员）的变体含有RNA 9片段，其末端序列与野生型病毒的RNA 9相同，但短约140个碱基对（bp）。体外翻译显示，野生型病毒的RNA 9产生次要结构蛋白VP 6（分子量38 K），而变体RNA 9编码32 K蛋白。对[32 P]-pCp标记的野生型和变体RNA 9分子的混合物进行解链和再退火后形成的杂合分子的分析表明，变体RNA 9的产生可能涉及在距离野生型RNA分子一端148 bp的位置处丢失约150 bp。在野生型和变体RNA 9的体外翻译期间通过提前终止产生的次要蛋白质的分析表明，缺失发生在野生型RNA 9的正链的3“末端。观察到蓝舌病病毒21型和Bunyip Creek（Palyam血清群成员）的RNA基因组片段10和9分别形成多联体。分子量估计和T1 RNase图谱表明，串联体是5“-3”至5“-3”方向的二聚体。二聚体RNA的体外翻译产生的产物与单体RNA产生的产物明显相同。这些结果与环状病毒进化的可能后果进行了讨论。
Orbivirus variants containing either RNA deletions or concatemeric RNAs have been isolated. A variant of Ibaraki virus (a member of the epizootic hemorrhagic disease of deer serogroup) contained an RNA 9 segment which had terminal sequences identical to RNA 9 of wild type virus but was approximately 140 base pairs (bp) shorter. In vitro translation showed that whereas RNA 9 of wild type virus generated the minor structural protein VP6 (molecular weight 38 K), the variant RNA 9 coded for a 32 K protein. Analysis of hybrid molecules formed after melting and reannealing mixtures of [32P]-pCp-labeled wild type and variant RNA 9 molecules indicated that generation of variant RNA 9 may have involved the loss of approximately 150 bp at a location 148 bp from one end of the wild type RNA molecule. Analysis of minor proteins generated by premature termination during in vitro translation of wild type and variant RNA 9 suggested that the deletion occurred towards the 3'' end of the positive strand of wild type RNA 9. RNA genome segments 10 and 9 of bluetongue virus type 21 and Bunyip Creek (a Palyam serogroup member) respectively, were observed to form concatemers. Molecular weight estimates and T1 RNase mapping suggested that the concatemers were dimers in a 5''-3'' to 5''-3'' orientation. In vitro translation of dimeric RNAs yielded products apparently identical to those generated by monomeric RNAs. The possible ramifications of these results with respect to orbivirus evolution are discussed.