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dc.contributor.authorHeighway, Jim
dc.contributor.authorGeurts van Kessel, A H
dc.date.accessioned2010-11-08T10:45:12Z
dc.date.available2010-11-08T10:45:12Z
dc.date.issued1987-04-24
dc.identifier.citationIsolation of a human genomic fragment, co-amplified with c-Ki-ras, that affects plasmid supercoiling in E. coli. 1987, 15 (8):3411-20 Nucleic Acids Res.en
dc.identifier.issn0305-1048
dc.identifier.pmid3033603
dc.identifier.doi10.1093/nar/15.8.3411
dc.identifier.urihttp://hdl.handle.net/10541/114928
dc.description.abstractAmplification of cellular proto-oncogenes has been implicated in the development of human malignancies. A library was constructed from genomic DNA extracted from a lung tumour, previously shown to carry an amplified c-Ki-ras 2 gene. Using a v-Ki-ras probe, a fragment with ras homology was isolated and shown to be amplified in the original tumour DNA to the same level as c-Ki-ras. Studies with human hamster hybrids demonstrated that it is normally located on human chromosome 12 (as is c-Ki-ras). The restriction map of the fragment is different from that of the known Ha, Ki or N-ras genes and its sequence shows evolutionary conservation, as demonstrated by hybridisation to the genomic DNA of several mammalian species. A pUC19 subclone (pK42), carrying a 1.3kb insert, shows supercoil heterogeneity in plasmid preparations, as does a second compatible plasmid introduced into the same bacterial host with pK42. It appears therefore that the subclone is encoding a product that affects DNA topoisomerase activity in E. coli.
dc.language.isoenen
dc.subjectCancer DNAen
dc.subjectLung Canceren
dc.subject.meshAnimals
dc.subject.meshBacterial Proteins
dc.subject.meshChromosome Mapping
dc.subject.meshChromosomes, Human, Pair 12
dc.subject.meshCricetinae
dc.subject.meshDNA Topoisomerases, Type I
dc.subject.meshDNA, Neoplasm
dc.subject.meshDNA, Superhelical
dc.subject.meshEscherichia coli
dc.subject.meshGene Amplification
dc.subject.meshHumans
dc.subject.meshHybrid Cells
dc.subject.meshLung Neoplasms
dc.subject.meshNucleotide Mapping
dc.subject.meshPlasmids
dc.subject.meshProto-Oncogene Proteins
dc.subject.meshProto-Oncogene Proteins p21(ras)
dc.titleIsolation of a human genomic fragment, co-amplified with c-Ki-ras, that affects plasmid supercoiling in E. coli.en
dc.typeArticleen
dc.identifier.journalNucleic Acids Researchen
html.description.abstractAmplification of cellular proto-oncogenes has been implicated in the development of human malignancies. A library was constructed from genomic DNA extracted from a lung tumour, previously shown to carry an amplified c-Ki-ras 2 gene. Using a v-Ki-ras probe, a fragment with ras homology was isolated and shown to be amplified in the original tumour DNA to the same level as c-Ki-ras. Studies with human hamster hybrids demonstrated that it is normally located on human chromosome 12 (as is c-Ki-ras). The restriction map of the fragment is different from that of the known Ha, Ki or N-ras genes and its sequence shows evolutionary conservation, as demonstrated by hybridisation to the genomic DNA of several mammalian species. A pUC19 subclone (pK42), carrying a 1.3kb insert, shows supercoil heterogeneity in plasmid preparations, as does a second compatible plasmid introduced into the same bacterial host with pK42. It appears therefore that the subclone is encoding a product that affects DNA topoisomerase activity in E. coli.


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