Chen D, Burns J W, Estes M K, Ramig R F
Division of Molecular Virology, Baylor College of Medicine, Houston, TX 77030.
Proc Natl Acad Sci U S A. 1989 May;86(10):3743-7. doi: 10.1073/pnas.86.10.3743.
We have previously characterized the biological and immunological properties of a simian rotavirus SA11 variant (4F) with an altered genome segment 4. The SA11-4F variant formed large plaques in the presence of protease, formed small clear plaques in the absence of protease, and grew to high titer in the presence of protease when compared to our standard wild type (SA11 clone 3). To determine the genome segment of the rotavirus SA11 variant 4F that encoded the unique protease-associated phenotypes of the variant, reassortants were generated that segregated the outer capsid genes of 4F onto a genetic background derived from either the bovine rotavirus B223 or our standard SA11 wild type (clone 3), both of which have contrasting protease-associated phenotypes. The parental and reassortant viruses were examined to determine which genes from the 4F variant encoded the ability (i) to form large plaques in the presence of protease, (ii) to form small clear plaques in the absence of exogenous protease, and (iii) to grow to significantly higher titer in the presence of protease. These phenotypes could be transferred to a clone 3 genetic background by a single genome segment from the 4F variant segment 4. However, in the 4F/B223 reassortants a different and unexpected situation was found. On a B223 genetic background the same phenotypes segregated with a combination of a minimum of two 4F genome segments, segments 4 and 9. These results indicate that the recipient genetic background onto which the genes of a donor rotavirus are reassorted can affect the phenotypes conferred by the presence of the donor segments. Thus, the results of segregation mapping experiments using reassortant viruses should be interpreted with caution.
我们之前已对一种基因组片段4发生改变的猿猴轮状病毒SA11变体(4F)的生物学和免疫学特性进行了表征。与我们的标准野生型(SA11克隆3)相比,SA11 - 4F变体在蛋白酶存在的情况下形成大的噬斑,在无蛋白酶的情况下形成小的清亮噬斑,并且在蛋白酶存在时能生长至高滴度。为了确定编码该变体独特蛋白酶相关表型的轮状病毒SA11变体4F的基因组片段,构建了重配病毒,将4F的外衣壳基因分离到源自牛轮状病毒B223或我们的标准SA11野生型(克隆3)的遗传背景上,这两种病毒具有截然不同的蛋白酶相关表型。对亲代病毒和重配病毒进行检测,以确定4F变体的哪些基因编码了以下能力:(i)在蛋白酶存在时形成大噬斑;(ii)在无外源性蛋白酶时形成小的清亮噬斑;(iii)在蛋白酶存在时生长至显著更高滴度。这些表型可通过4F变体片段4的单个基因组片段转移到克隆3的遗传背景上。然而,在4F/B223重配病毒中发现了一种不同且意想不到的情况。在B223遗传背景下,相同的表型与至少两个4F基因组片段(片段4和9)的组合相关联。这些结果表明,供体轮状病毒基因重配到的受体遗传背景可影响供体片段存在所赋予的表型。因此,使用重配病毒进行的分离图谱实验结果应谨慎解读。