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941.
Depurination-induced infidelity of deoxyribonucleic acid synthesis with purified deoxyribonucleic acid replication proteins in vitro 总被引:18,自引:0,他引:18
Removal of purine bases from phi X174 single-stranded DNA leads to increased reversion frequency of amber mutations when this DNA is copied in vitro with purified DNA polymerases. This depurination-induced mutagenesis is observed at three different genetic loci and with several different purified enzymes, including Escherichia coli DNA polymerases I and III, avian myeloblastosis virus DNA polymerase, and eukaryotic DNA polymerases alpha, beta, and gamma. The extent of mutagenesis correlates with the estimated frequency of bypass of the lesion and is greatest with inherently inaccurate DNA polymerases which lack proofreading capacity. With E. coli DNA polymerase I, conditions which diminish proofreading result in a 3-5-fold increase in depurination-induced mutagenesis, suggesting a role for proofreading in determining the frequency of bypass of apurinic sites. The addition of E. coli single-stranded DNA-binding protein to polymerase I catalyzed reactions with depurinated DNA had no effect on the extent of mutagenesis. Analysis of wild-type revertants produced during in vitro DNA synthesis by polymerase I or avian myeloblastosis virus DNA polymerase on depurinated phi X174 amber 3 DNA indicates a preference for insertion of dAMP opposite the putative apurinic site at position 587. These results are discussed in relation both to the mutagenic potential of apurinic sites in higher organisms and to studies on error-prone DNA synthesis. 相似文献
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The clinical and laboratory findings in seven children with Kawasaki disease are reviewed. Four of the patients had the more complicated course that has characterized the cases diagnosed in North America. This suggests that the benign forms are often mistaken for other febrile illnesses. The patients were two girls and five boys ranging in age from 4 months to 7 years; six were Caucasian and one was a North American Indian. Fever, redness of the oral mucosa, an erythematous or scarlatiniform rash and cervical adenopathy were seen in all; six patients had the characteristic fingertip desquamation and nonexudative conjunctivitis. Cardiac involvement occurred in four patients, two of whom had coronary artery aneurysm or thrombosis. Arthritis or arthralgia was seen in six patients, and aseptic meningitis occurred in four. Of the three patients with jaundice two underwent laparotomy and excision of a hydropic gallbladder; one of them died from Klebsiella pneumoniae sepsis and disseminated intravascular coagulopathy. 相似文献
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Aliphatic aldehydes of different chain lengths were found to differ in their reaction at 22 °C with the B. harveyi luciferase peroxyflavin intermediate. Although similar quantum yields were obtained in the luciferase reaction with the different chain-length aldehydes, the catalytic turnover rates differed. The kinetics of a reaction utilizing two aldehydes of different chain lengths can thus indicate the degree to which the aldehyde reaction is reversible. By such criteria the reactions of octanal and decanal were found to be readily reversible, while that of dodecanal was not. This conclusion was supported both by the effects of long-chain alcohols, which are competitive inhibitors, and by the secondary addition of hydroxylamine, an aldehyde trapping agent. The results are consistent with a model in which there are many intermediates along the reaction path. Since the reactions are monitored by decay of luminescence intensity, it is difficult to determine the position of the rate-determining step. For octanal and decanal the rate-limiting step could be at an early reversible stage of the reaction, but later for dodecanal, subsequent to a less reversible step, but still prior to the final irreversible step which populates the excited state. 相似文献
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