Taketo A, Kodaira K
Biochim Biophys Acta. 1978 Oct 24;520(3):505-11. doi: 10.1016/0005-2787(78)90136-3.
Host functions required for replication of microvirid phage G13 DNA were investigated in vivo, using thermosensitive dna mutants of Escherichia coli. In dna+ bacteria, conversion of viral single-stranded DNA into double-stranded replicative form (stage I synthesis) was resistant to 150 microgram/ml of chloramphenicol or 200 microgram/ml of rifampicin. Although multiplication of G13 phage was severely inhibited at 42--43 degrees C even in dna+ host, considerable amount of parental replicative form was synthesized at 43 degrees C in dna+, dnaA or dnaE bacteria. In dnaB and dnaG mutants, however, synthesis of parental replicative form was severely inhibited at the restrictive temperature. Interestingly enough, stage I replication of G13 DNA was, unlike that of phiX174, dependent on host dnaC(D) function. Moreover, the stage I synthesis of G13 DNA in dnaZ was thermosensitive in nutrient broth but not in Tris/casamino acids/glucose medium. In contrast with the stage I replication, synthesis of G13 progeny replicative form was remarkably thermosensitive even in dna+ or dnA cells.
利用大肠杆菌的温度敏感型dna突变体,在体内研究了微小病毒噬菌体G13 DNA复制所需的宿主功能。在dna⁺细菌中,病毒单链DNA转化为双链复制型(第一阶段合成)对150微克/毫升氯霉素或200微克/毫升利福平具有抗性。尽管即使在dna⁺宿主中,G13噬菌体的增殖在42 - 43℃时也受到严重抑制,但在dna⁺、dnaA或dnaE细菌中,在43℃时仍合成了相当数量的亲本复制型。然而,在dnaB和dnaG突变体中,在限制温度下亲本复制型的合成受到严重抑制。有趣的是,与phiX174不同,G13 DNA的第一阶段复制依赖于宿主dnaC(D)功能。此外,G13 DNA在dnaZ中的第一阶段合成在营养肉汤中对温度敏感,但在Tris/酪蛋白氨基酸/葡萄糖培养基中则不敏感。与第一阶段复制相反,即使在dna⁺或dnA细胞中,G13子代复制型的合成对温度也非常敏感。