Pacumbaba R P, Center M S
J Virol. 1974 Dec;14(6):1380-7. doi: 10.1128/JVI.14.6.1380-1387.1974.
Infection of Escherichia coli with bacteriophage T7 results in the formation of an endonuclease which is selectively associated with the T7 DNA-membrane complex. A specificity of association with the complex is indicated by the finding that the enzyme is completely resolved from a previously described T7 endonuclease I. When membrane complexes containing (3)H-labeled in vivo synthesized DNA are incubated in the standard reaction mixture a specific cleavage product is formed which is about one-fourth the size of T7 DNA. The endonuclease associated with the complex produces a similar cleavage product after extensive incubation with native T7 DNA or T7 concatemers. Degradation of concatemers occurs by a mechanism in which the DNA is converted to molecules one-half the size of T7. This product is in turn converted to fragments one-fourth the size of mature phage DNA. The endonuclease is not present in membrane complexes from uninfected cells or cells infected with gene 1 mutants. The enzyme activity is, however, present in cells infected with mutants defective in T7 DNA synthesis or maturation.
用噬菌体T7感染大肠杆菌会导致一种核酸内切酶的形成,该酶与T7 DNA - 膜复合物选择性相关。与该复合物结合的特异性体现在这一发现上:该酶与先前描述的T7核酸内切酶I完全不同。当含有体内合成的(3)H标记DNA的膜复合物在标准反应混合物中孵育时,会形成一种特定的切割产物,其大小约为T7 DNA的四分之一。与该复合物相关的核酸内切酶在与天然T7 DNA或T7多联体广泛孵育后会产生类似的切割产物。多联体的降解通过一种机制发生,其中DNA被转化为大小为T7一半的分子。该产物又会被转化为成熟噬菌体DNA四分之一大小的片段。未感染细胞或感染基因1突变体的细胞的膜复合物中不存在这种核酸内切酶。然而,该酶活性存在于感染了T7 DNA合成或成熟有缺陷的突变体的细胞中。