在大肠杆菌提取物中将噬菌体φX174和fd单链DNA转化为复制形式。
Conversion of phiX174 and fd single-stranded DNA to replicative forms in extracts of Escherichia coli.
作者信息
Wickner R B, Wright M, Wickner S, Hurwitz J
出版信息
Proc Natl Acad Sci U S A. 1972 Nov;69(11):3233-7. doi: 10.1073/pnas.69.11.3233.
varphiX174 and M13 (fd) single-stranded circular DNAs are converted to their replicative forms by extracts of E. coli pol A1 cells. We find that the varphiX174 DNA-dependent reaction requires Mg(++), ATP, and all four deoxynucleoside triphosphates, but not CTP, UTP, or GTP. This reaction also involves the products of the dnaC, dnaD, dnaE (DNA polymerase III), and dnaG genes, but not that of dnaF (ribonucleotide reductase). The in vitro conversion of fd single-stranded DNA to the replicative form requires all four ribonucleoside triphosphates, Mg(++), and all four deoxynucleoside triphosphates. The reaction involves the product of gene dnaE but not those of genes dnaC, dnaD, dnaF, or dnaG. The reaction with fd DNA is inhibited by rifampicin or antibody to RNA polymerase, while the reaction with varphiX174 DNA is not affected by either. With the varphiX174 DNA-dependent reaction, activities have been detected that specifically complement extracts of dnaA, dnaB, dnaC, dnaD, or dnaG mutants.
φX174和M13（fd）单链环状DNA可被大肠杆菌pol A1细胞提取物转化为它们的复制形式。我们发现，φX174 DNA依赖性反应需要Mg（++）、ATP和所有四种脱氧核苷三磷酸，但不需要CTP、UTP或GTP。该反应还涉及dnaC、dnaD、dnaE（DNA聚合酶III）和dnaG基因的产物，但不涉及dnaF（核糖核苷酸还原酶）的产物。fd单链DNA体外转化为复制形式需要所有四种核糖核苷三磷酸、Mg（++）和所有四种脱氧核苷三磷酸。该反应涉及dnaE基因的产物，但不涉及dnaC、dnaD、dnaF或dnaG基因的产物。与fd DNA的反应受到利福平或RNA聚合酶抗体的抑制，而与φX174 DNA的反应则不受二者影响。对于φX174 DNA依赖性反应，已检测到能特异性互补dnaA、dnaB、dnaC、dnaD或dnaG突变体提取物的活性。
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