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  • 1
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    Genomic sequencing and methylation analysis by ligation mediated PCR (1989)
    American Association for the Advancement of Science (AAAS)
    Details
    Publication Date: 1989-11-10
    Description: Genomic sequencing permits studies of in vivo DNA methylation and protein-DNA interactions, but its use has been limited because of the complexity of the mammalian genome. A newly developed genomic sequencing procedure in which a ligation mediated polymerase chain reaction (PCR) is used generates high quality, reproducible sequence ladders starting with only 1 microgram of uncloned mammalian DNA per reaction. Different sequence ladders can be created simultaneously by inclusion of multiple primers and visualized separately by rehybridization. Relatively little radioactivity is needed for hybridization and exposure times are short. Methylation patterns in genomic DNA are readily detectable; for example, 17 CpG dinucleotides in the 5' region of human X-linked PGK-1 (phosphoglycerate kinase 1) were found to be methylated on an inactive human X chromosome, but unmethylated on an active X chromosome.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Pfeifer, G P -- Steigerwald, S D -- Mueller, P R -- Wold, B -- Riggs, A D -- AG08196/AG/NIA NIH HHS/ -- GM355262BW/GM/NIGMS NIH HHS/ -- RR07003/RR/NCRR NIH HHS/ -- New York, N.Y. -- Science. 1989 Nov 10;246(4931):810-3.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Molecular Biology Section, Beckman Research Institute of the City of Hope, Duarte, CA 91010.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/2814502" target="_blank"〉PubMed〈/a〉
    Keywords: 5-Methylcytosine ; Animals ; Autoradiography ; Base Sequence ; Cytosine ; DNA/*genetics/metabolism ; Exons ; HeLa Cells ; Humans ; Methylation ; Molecular Sequence Data ; *Nucleic Acid Amplification Techniques ; *Nucleic Acid Hybridization ; Phosphoglycerate Kinase/genetics ; Polymerase Chain Reaction/*methods ; Promoter Regions, Genetic ; X Chromosome
    Print ISSN: 0036-8075
    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Computer Science , Medicine , Natural Sciences in General , Physics
    Published by American Association for the Advancement of Science (AAAS)
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  • 2
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    Preferential formation of benzo[a]pyrene adducts at lung cancer mutational hotspots in P53 (1996)
    American Association for the Advancement of Science (AAAS)
    Details
    Publication Date: 1996-10-18
    Description: Cigarette smoke carcinogens such as benzo[a]pyrene are implicated in the development of lung cancer. The distribution of benzo[a]pyrene diol epoxide (BPDE) adducts along exons of the P53 gene in BPDE-treated HeLa cells and bronchial epithelial cells was mapped at nucleotide resolution. Strong and selective adduct formation occurred at guanine positions in codons 157, 248, and 273. These same positions are the major mutational hotspots in human lung cancers. Thus, targeted adduct formation rather than phenotypic selection appears to shape the P53 mutational spectrum in lung cancer. These results provide a direct etiological link between a defined chemical carcinogen and human cancer.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Denissenko, M F -- Pao, A -- Tang, M -- Pfeifer, G P -- CA65652/CA/NCI NIH HHS/ -- ES03124/ES/NIEHS NIH HHS/ -- New York, N.Y. -- Science. 1996 Oct 18;274(5286):430-2.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Department of Biology, Beckman Research Institute of the City of Hope, Duarte, CA 91010, USA.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/8832894" target="_blank"〉PubMed〈/a〉
    Keywords: 7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide/metabolism/*toxicity ; Bronchi ; Carcinogens/metabolism/*toxicity ; Cells, Cultured ; Codon ; DNA Adducts/*metabolism ; Dinucleoside Phosphates/metabolism ; Exons ; Fibroblasts ; *Genes, p53 ; HeLa Cells ; Humans ; Lung Neoplasms/etiology/*genetics ; Mutagens/metabolism/toxicity ; *Mutation ; Plants, Toxic ; Smoke/adverse effects ; Tobacco
    Print ISSN: 0036-8075
    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Computer Science , Medicine , Natural Sciences in General , Physics
    Published by American Association for the Advancement of Science (AAAS)
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  • 3
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    Slow repair of pyrimidine dimers at p53 mutation hotspots in skin cancer (1994)
    American Association for the Advancement of Science (AAAS)
    Details
    Publication Date: 1994-03-11
    Description: Ultraviolet light has been linked with the development of human skin cancers. Such cancers often exhibit mutations in the p53 tumor suppressor gene. Ligation-mediated polymerase chain reaction was used to analyze at nucleotide resolution the repair of cyclobutane pyrimidine dimers along the p53 gene in ultraviolet-irradiated human fibroblasts. Repair rates at individual nucleotides were highly variable and sequence-dependent. Slow repair was seen at seven of eight positions frequently mutated in skin cancer, suggesting that repair efficiency may strongly contribute to the mutation spectrum in a cancer-associated gene.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Tornaletti, S -- Pfeifer, G P -- ES06070/ES/NIEHS NIH HHS/ -- New York, N.Y. -- Science. 1994 Mar 11;263(5152):1436-8.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Beckman Rsearch Institute of the City of Hope, Department of Biology, Duarte, CA 91010.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/8128225" target="_blank"〉PubMed〈/a〉
    Keywords: Cells, Cultured ; *DNA Repair ; Exons ; *Genes, p53 ; HeLa Cells ; Humans ; Mutation ; Phosphoglycerate Kinase/genetics ; Polymerase Chain Reaction ; Pyrimidine Dimers/*metabolism ; Skin/metabolism/*radiation effects ; Skin Neoplasms/*genetics/metabolism ; Ultraviolet Rays
    Print ISSN: 0036-8075
    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Computer Science , Medicine , Natural Sciences in General , Physics
    Published by American Association for the Advancement of Science (AAAS)
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