PHYSICAL MAPPING AND DNA-SEQUENCE ANALYSIS OF THE RIFAMPICIN RESISTANCE LOCUS IN VACCINIA VIRUS
PHYSICAL MAPPING AND DNA-SEQUENCE ANALYSIS OF THE RIFAMPICIN RESISTANCE LOCUS IN VACCINIA VIRUS
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DOI：
10.1016/0042-6822(85)90141-2
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发表时间：
1985-01-01
期刊：
影响因子：
3.7
通讯作者：
PAOLETTI, E
中科院分区：
文献类型：
--
作者：
TARTAGLIA, J;PAOLETTI, E
利福平已被证明在包膜形成的离散步骤中抑制痘病毒的成熟(Moss等人，1969；Pennington等人，1970；Nagayama等人，1970；Grimley等人，1970)。选择了一个抗利福平的痘苗病毒突变体(RIFR)，因为它能在100微克/毫升的利福平存在下生长。利用来自RIFR突变病毒的完整DNA或限制性内切酶克隆的DNA亚片段，通过标记拯救分析，在HindIII D片段内唯一的XhoI位点的左侧物理定位了利福平抗性的基因座。对包含该区域的445bp片段进行DNA测序，与相应的野生型DNA片段相比，显示了AT到GC的转变。对445bp片段中6个潜在开放阅读框的分析表明，只有一个可用开放阅读框。在此基础上，对利福平耐药痘苗病毒突变株进行了从天冬酰胺到天冬氨酸的密码子转换。
Rifampicin has been shown to inhibit the maturation of poxviruses at a discrete step in envelope formation (Moss et al., 1969; Pennington et al., 1970; Nagayama et al., 1970; Grimley et al., 1970). A rifampicin-resistant vaccinia virus mutant (RifR) was selected for its ability to grow in the presence of 100 .mu.g/ml of rifampicin. Utilizing intact DNA or endonuclease restricted cloned DNA subfragments derived from the RifR mutant virus, the locus specifying rifampicin resistance was physically mapped by marker rescue analysis leftward of the unique XhoI site within the HindIII D fragment. DNA sequencing of a 445 bp fragment encompassing this region revealed an AT to GC transition when compared with the equivalent wild-type DNA fragment. Analysis of the six potential open reading frames within the 445-bp fragment indicated only one available open reading frame. On this basis, the rifampicin-resistant vaccinia virus mutant was shown to have a codon transition from asparagine to aspartic acid.