Molecular genetics of herpes simplex virus. VII. Characterization of a temperature-sensitive mutant produced by in vitro mutagenesis and defective in DNA synthesis and accumulation of gamma polypeptides
Molecular genetics of herpes simplex virus. VII. Characterization of a temperature-sensitive mutant produced by in vitro mutagenesis and defective in DNA synthesis and accumulation of gamma polypeptides
复制标题
单纯疱疹病毒的分子遗传学。
作者：
A. Conley;D. Knipe;P. Jones;B. Roizman
我们报告的性质的温度敏感的突变体产生的转染细胞与完整的DNA和特定的DNA片段诱变低水平的羟胺。突变体在39 ℃下的平板接种效率相对于在33.5 ℃下的平板接种效率为5 × 10 ~（-6）。在非允许温度下产生的多肽的模式与用抑制浓度的膦酰基乙酸处理的感染细胞中的野生型病毒所见的相似，因为产生了α和β多肽，而大多数γ多肽减少或不存在。一致地，突变体不产生病毒DNA，尽管不能证明病毒DNA聚合酶的温度敏感性。用单纯疱疹病毒2型（HSV-2）DNA进行的标记拯救研究将L组分中的突变体定位在HSV-1基因组原型（P）排列的图谱位置0.385和0.402内。重组体的分析允许对指定感染细胞多肽36、35、37、19.5、11、8、2、43和44的基因进行定位，但只有HSV-2的感染细胞多肽8由所有含有可证实的HSV-2序列的重组体一致地产生。用克隆的HSV-1 DNA片段进行的标记拯救研究在0.383和0.388图谱单位之间的小于10（3）个碱基对内绘制了温度敏感性病变。与克隆的HSV-1 DNA杂交的RNA的翻译，包括含有突变的最小区域，揭示了多肽8（128，000分子量），其先前被鉴定为对病毒DNA具有高亲和力的β多肽，以及先前在标记细胞的裂解物中未鉴定的多肽（25，000分子量）。
We report on the properties of a temperature-sensitive mutant produced by transfection of cells with intact DNA and a specific DNA fragment mutagenized with low levels of hydroxylamine. The plating efficiency of the mutant at 39 degrees C relative to that at 33.5 degrees C was 5 X 10(-6). The pattern of polypeptides produced at the nonpermissive temperature was similar to that seen with wild-type virus in infected cells treated with inhibitory concentrations of phosphonoacetic acid in that alpha and beta polypeptides were produced, whereas most gamma polypeptides were either reduced or absent. Consistently, the mutant did not make viral DNA, although temperature sensitivity of the viral DNA polymerase could not be demonstrated. Marker rescue studies with herpes simplex virus type 2 (HSV-2) DNA mapped the mutant in the L component within map positions 0.385 and 0.402 in the prototype (P) arrangement of the HSV-1 genome. Analysis of the recombinants permitted the mapping of the genes specifying infected cell polypeptides 36, 35, 37, 19.5, 11, 8, 2, 43, and 44, but only the infected cell polypeptide 8 of HSV-2 was consistently made by all recombinants containing demonstrable HSV-2 sequences. Marker rescue studies with cloned HSV-1 DNA fragments mapped the temperature-sensitive lesion within less than 10(3) base pairs between 0.383 and 0.388 map units. Translation of the RNA hybridizing to cloned HSV-1 DNA, encompassing the smallest region containing the mutation, revealed polypeptide 8 (128,000 molecular weight), which was previously identified as a beta polypeptide with high affinity for viral DNA, and a polypeptide (25,000 molecular weight) not previously identified in lysates of labeled cells.
将重复和非重复单纯疱疹病毒 1 序列克隆为 BamHI 片段。
DOI：
10.1073/pnas.77.7.4201
发表时间：
1980
影响因子：
11.1
作者：
Post,LE;Conley,AJ;Mocarski,ES;Roizman,B
通讯作者：
Roizman,B