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1.
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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We are investigating the mechanisms for producing or avoiding errors during DNA synthesis catalyzed by DNA replication and repair proteins purified from eukaryotic sources. Using assays that monitor the fidelity of a single round of DNA synthesis in vitro, we have defined the error frequency and mutational specificity of the four classes of animal cell DNA polymerases (alpha, beta, delta, gamma), and the fidelity of an SV40 origin-dependent DNA replication complex in extracts of HeLa cells. 相似文献
4.
肺癌在中国恶性肿瘤的发病率位居第一,随着低剂量薄层CT在肺癌筛查中的广泛应用,临床发现更多表现为非完全实性结节的肺腺癌,目前众多研究使CT影像学特征和肺腺癌病理的关系得到更进一步的认知,虽然CT能对部分非完全实性结节做出定性和定位诊断,但仍有部分非完全实性结节诊断困难,PET-CT结合了病灶的代谢信息和精确的定位信息,从而提高对肺部结节诊断的敏感性、特异性、准确性,综合多个文献PET-CT在非完全实性结节中的诊断分期价值较CT无明显提升,却在评估预后和制定合适手术方案上可以起到一定的作用,本文就PET-CT在SSN中的应用价值进行阐述。 相似文献
5.
Douglas S. Richmond† Brian A. Kunkel Nethi Somasekhar Parwinder S. Grewal 《Ecological Entomology》2004,29(3):353-360
Abstract. 1. The fungus Neotyphodium lolii forms a symbiotic relationship with its grass host Lolium perenne (perennial ryegrass). The fungus benefits from access to plant nutrients and photosynthate, whereas the plant benefits from acquired chemical defence against herbivory.
2. This study examined the potential for endophyte-mediated plant defences to influence interactions between fall armyworm Spodoptera frugiperda , and the entomopathogenic nematode Steinernema carpocapsae and clarified biological mechanisms underlying the observations made.
3. In laboratory and greenhouse experiments, S. frugiperda larvae were fed endophytic or non-endophytic L. perenne then exposed to S. carpocapsae or injected with the nematodes' symbiotic bacteria Xenorhabdus nematophila .
4. In all instances, S. frugiperda larvae fed endophyte-infected grass suffered significantly lower mortality than those fed non-endophytic plants. Although larvae fed endophyte-infected grass often had significantly lower biomass than those fed uninfected grass, these differences did not account for altered susceptibility to S. carpocapsae .
5. Endophyte-mediated reductions in herbivore susceptibility to the nematode pathogen represent a herbivore adaptation that effectively turns the tables on both plant and natural enemy by reducing the virulence of the nematodes' symbiotic bacteria while expanding the temporal window of herbivory. 相似文献
2. This study examined the potential for endophyte-mediated plant defences to influence interactions between fall armyworm Spodoptera frugiperda , and the entomopathogenic nematode Steinernema carpocapsae and clarified biological mechanisms underlying the observations made.
3. In laboratory and greenhouse experiments, S. frugiperda larvae were fed endophytic or non-endophytic L. perenne then exposed to S. carpocapsae or injected with the nematodes' symbiotic bacteria Xenorhabdus nematophila .
4. In all instances, S. frugiperda larvae fed endophyte-infected grass suffered significantly lower mortality than those fed non-endophytic plants. Although larvae fed endophyte-infected grass often had significantly lower biomass than those fed uninfected grass, these differences did not account for altered susceptibility to S. carpocapsae .
5. Endophyte-mediated reductions in herbivore susceptibility to the nematode pathogen represent a herbivore adaptation that effectively turns the tables on both plant and natural enemy by reducing the virulence of the nematodes' symbiotic bacteria while expanding the temporal window of herbivory. 相似文献
6.
Localization and cloning of Xp21 deletion breakpoints involved in muscular dystrophy 总被引:26,自引:2,他引:24
Anthony P. Monaco Corlee J. Bertelson Chris Colletti-Feener Louis M. Kunkel 《Human genetics》1987,75(3):221-227
Summary Twenty-nine deletion breakpoints were mapped in 220 kb of the DXS164 locus relative to potential exons of the Duchenne and Becker muscular dystrophy gene. Four deletion junction fragments were isolated to acquire outlying Xp21 loci on both the terminal and centromere side of the DXS164 locus. The junction loci were used for chromosome walking, searches for DNA polymorphisms, and mapping against deletion and translocation breakpoints. Forty-four unrelated deletions were analyzed using the junction loci as hybridization probes to map the endpoints between cloned Xp21 loci. DNA polymorphisms from the DXS164 and junction loci were used to follow the segregation of a mutation in a family that represents a recombinant. Both the physical and genetic data point to a very large size for this X-linked muscular dystrophy locus. 相似文献
7.
The beta form is the dominant interleukin 1 released by murine peritoneal macrophages 总被引:3,自引:0,他引:3
S W Chensue C Shmyr-Forsch I G Otterness S L Kunkel 《Biochemical and biophysical research communications》1989,160(1):404-408
Using highly specific polyclonal antisera raised against recombinant murine IL-1 alpha and beta, we performed solid-phase immunoabsorption studies on supernates of resident and adjuvant-elicited CBA/J mouse peritoneal macrophages. Antibody specificity was established by reciprocal absorption studies and Western blot analysis. Supernates obtained from macrophages cultured for 18 hr in the presence of 1 microgram/ml lipopolysaccharide (LPS) were subjected to immunoabsorption. Approximately 78-90% of the released bioactive material was IL-1 and about 80% of this could be attributed to IL-1 beta. Analogous to that reported for human monocytes, these data suggest that IL-1 beta is the predominant released form of IL-1. 相似文献
8.
The aim of this study was to investigate circadian variation in concentrations of arachidonic acid (AA) metabolites in relation to the circadian pattern in bronchial patency. Blood samples were obtained at 4-hr intervals from 2000 of 1 day until 1400 of the next from 12 diurnally active asthmatic and six diurnally active non-asthmatic patients. Bloods were analyzed for the prostanoids thromboxane A2 (measured as stable metabolite 6-keto-PGF1a), PGE2 and PGF2a. Airways patency was assessed by self-measurement of peak expiratory flow (PEF). In asthmatics, circadian variation was detected in PEF as well as PGE2 and TXB2. The circadian trough of the PEF rhythm closely coincided with the circadian peak of the PGE2 and TXB2 rhythms. In the controls, the PEF was not circadian rhythmic. Of the AA metabolites only 6-keto-PGF1a exhibited 24-hr bioperiodicity in the controls. The controls exhibited a significantly higher circadian mean of PEF (P less than 0.001), while the asthmatics had a lower 24-hr average PGE2 but greater mean TXB2/PGE2 ratio. The obstructive effect caused by the overall 24-hr deficiency of PGE2 in asthmatics is possibly amplified by the increased of TXB2 during the early morning hours. This dissociation of the temporal patterns in TXB2 and PGE2 levels over the 24 hr is discussed as a characteristic finding for asthmatics. 相似文献
9.
A 10-megabase physical map of human Xp21, including the Duchenne muscular dystrophy gene 总被引:26,自引:0,他引:26
M Burmeister A P Monaco E F Gillard G J van Ommen N A Affara M A Ferguson-Smith L M Kunkel H Lehrach 《Genomics》1988,2(3):189-202
Using pulsed-field gel electrophoresis and 12 Xp21-derived DNA probes, we have constructed a continuous restriction map spanning more than 4 million base pairs (4 Mbp), including the Duchenne muscular dystrophy gene of more than 2 Mbp. This detailed map is part of a less detailed map spanning 10 Mbp, also spanning the genes for glycerol kinase and congenital adrenal hypoplasia, constructed under electrophoresis conditions which separated DNA fragments in the range 200 to 4000 kbp. DNA from three different tissues was analyzed, and differential methylation was observed. 相似文献
10.
An explanation for the phenotypic differences between patients bearing partial deletions of the DMD locus 总被引:78,自引:0,他引:78
Deletions giving rise to Duchenne muscular dystrophy (DMD) and the less severe Becker muscular dystrophy (BMD) occur in the same large gene on the short arm of the human X chromosome. We present a molecular mechanism to explain the clinical difference in severity between DMD and BMD patients who bear partial deletions of the same gene locus. The model is based on the breakpoints of intragenic deletions and their effect on the translation of triplet codons into amino acids of the protein product. Deletions identified in three DMD patients are shown to shift the translational open reading frame (ORF) of triplet codons for amino acids, and each deletion is predicted to result in a truncated, abnormal protein product. Deletions identified in three BMD patients are shown to maintain the translational ORF for amino acids and predict a shorter, lower molecular weight protein. The smaller protein product is presumed to be semifunctional and to result in a milder clinical phenotype. The same ORF mechanism is also applicable to potential 5' and 3' intron splice mutations and their effect on protein production and clinical phenotype. 相似文献