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首页> 外文期刊>Journal of Virology >Marker rescue of temperature-sensitive mutations of vaccinia virus WR: correlation of genetic and physical maps.
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Marker rescue of temperature-sensitive mutations of vaccinia virus WR: correlation of genetic and physical maps.

机译：痘苗病毒温度敏感突变的标记拯救：遗传与物理图谱的相关性。

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The physical map locations of 62 temperature-sensitive mutations of vaccinia virus WR have been determined by marker rescue experiments, using cloned HindIII fragments of wild-type DNA. Since vaccinia virus DNA is not infectious, marker rescue was performed by infecting monolayers of cells at the nonpermissive temperature with a low multiplicity of the mutant to be rescued and transfecting with calcium phosphate-precipitated recombinant DNA. Wild-type recombinants were measured by using either a direct plaque assay technique or a two-step procedure in which the final yield of virus from the transfected cells was assayed at the permissive and nonpermissive temperatures. Mutants that had been previously assigned to the same complementation-recombination group were rescued by the same HindIII fragment, with the exception of three mutants in one group that were rescued by either one of two adjacent fragments. A comparison between the genetic linkage map of the temperature-sensitive mutations in 30 mutants with their physical locations demonstrated that not only was the order of the genetic map correct but also recombination frequencies generally reflected actual physical distances.
机译：使用克隆的HindIII片段的野生型DNA的标记救援实验确定了62个温度敏感突变的物理映射位置。由于疫苗病毒DNA不感染，因此通过在非智能温度下感染细胞单层细胞来进行标记物救援，从多种突变体拯救并用磷酸钙沉淀的重组DNA转染。通过使用直接斑块测定技术或两步程序测量野生型重组剂，其中在允许和非智能温度下测定从转染的细胞中的病毒的最终产量。先前分配给相同互动重组基团的突变体被相同的HindIII片段寄救，除了一个由两个相邻碎片中的一个抵押的一个组中的三个突变体。在30个突变体中的温度敏感突变的遗传联系地图与物理位置的比较证明，不仅是遗传图谱的顺序，而且还有重组频率通常反射实际的物理距离。

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