Zograf Iu N, Ogryz'ko V V, Bass I A, Chernyĭ D I
Mol Biol (Mosk). 1985 May-Jun;19(3):818-32.
The EcoRI fragment of T4 DNA containing the W-29 genes and its subfragments were cloned in the pBR322 and the singlestranded M13 phage. Hybridization with a cloned DNA showed that in T4 infected cells the transcription of the late genes 25-29 depends on the phage-induced RNA polymerase changes and on replication of phage DNA. At a late infection stage one also observes an enhanced transcription of the (early) genes uvsW and uvsY, which depends on viral DNA replication. Both early and late genes within recombinant plasmids are also expressed in uninfected cells carrying a plasmid regardless of the inserted fragment orientation and independently of the vector promoters. Hybrid plasmids demonstrated a high frequency of recombination with phage DNA in the infected cell. An RNA polymerase from uninfected cells binds itself to the late cloned genes to form "open" complexes. A purified RNA polymerase transcribes both early and late genes within recombinant plasmids. The relative transcription of the late cloned genes is enhanced if one uses an RNA polymerase from T4-infected cells. The super-helicity of template DNA is essential for transcription of early and late genes.
含有W - 29基因的T4 DNA的EcoRI片段及其亚片段被克隆到pBR322和单链M13噬菌体中。与克隆DNA的杂交表明,在T4感染的细胞中,晚期基因25 - 29的转录依赖于噬菌体诱导的RNA聚合酶变化以及噬菌体DNA的复制。在感染后期,还观察到(早期)基因uvsW和uvsY的转录增强,这依赖于病毒DNA复制。重组质粒中的早期和晚期基因在携带质粒的未感染细胞中也能表达,而与插入片段的方向无关,且不依赖于载体启动子。杂交质粒在感染细胞中与噬菌体DNA表现出高频率的重组。来自未感染细胞的RNA聚合酶自身结合到晚期克隆基因上形成“开放”复合物。纯化的RNA聚合酶转录重组质粒中的早期和晚期基因。如果使用来自T4感染细胞的RNA聚合酶,晚期克隆基因的相对转录会增强。模板DNA的超螺旋结构对于早期和晚期基因的转录至关重要。