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噬菌体T4新uvsW等位基因的分离与遗传特征分析

Isolation and genetic characterization of new uvsW alleles of bacteriophage T4.

作者信息

Derr L K, Drake J W

机构信息

Laboratory of Molecular Genetics, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709.

出版信息

Mol Gen Genet. 1990 Jul;222(2-3):257-64. doi: 10.1007/BF00633826.

Abstract

The uvsW gene of bacteriophage T4 is required for wild-type levels of recombination, for normal survival and mutagenesis after UV irradiation, and for wild-type resistance to hydroxyurea. Additionally, uvsW mutations restore the arrested DNA synthesis caused by mutations in any of several genes that block secondary initiation (recombination-primed replication, the major mode of initiation at late times), but only partially restore the reduced burst size. A uvsW deletion mutation was constructed to establish the null-allele phenotype, which is similar but not identical to the phenotype of the canonical uvsW mutation, and to demonstrate convincingly that the uvsW gene is nonessential (although uvsW mutations severely compromise phage production). In an attempt to uncouple the diverse effects of uvsW mutations, temperature-sensitive uvsWts mutants were isolated. Recombination and replication effects were partially uncoupled in these mutants, suggesting distinct and separable roles for uvsW in the two processes. Furthermore, the restoration of DNA synthesis but not recombination in the double mutants uvsW uvsX and uvsW uvsY prompts the hypothesis that the restored DNA synthesis is not recombinationally initiated.

摘要

噬菌体T4的uvsW基因对于野生型水平的重组、紫外线照射后的正常存活和诱变以及对羟基脲的野生型抗性是必需的。此外,uvsW突变可恢复由多个阻断二次起始(重组引发的复制,后期主要的起始模式)的基因中的任何一个突变所导致的DNA合成停滞,但只能部分恢复减少的爆发量。构建了一个uvsW缺失突变以确定无效等位基因表型,该表型与典型uvsW突变的表型相似但不完全相同,并令人信服地证明uvsW基因不是必需的(尽管uvsW突变严重损害噬菌体的产生)。为了分离uvsW突变的不同效应,分离了温度敏感型uvsWts突变体。在这些突变体中,重组和复制效应部分解偶联,表明uvsW在这两个过程中具有不同且可分离的作用。此外,uvsW uvsX和uvsW uvsY双突变体中DNA合成的恢复而非重组的恢复提示了这样一种假说,即恢复的DNA合成不是由重组引发的。

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