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dc.contributor.authorJelinek, J
dc.contributor.authorKleibl, K
dc.contributor.authorDexter, T Michael
dc.contributor.authorMargison, Geoffrey P
dc.date.accessioned2010-11-10T10:42:18Z
dc.date.available2010-11-10T10:42:18Z
dc.date.issued1988-01
dc.identifier.citationTransfection of murine multi-potent haemopoietic stem cells with an E. coli DNA alkyltransferase gene confers resistance to the toxic effects of alkylating agents. 1988, 9 (1):81-7 Carcinogenesisen
dc.identifier.issn0143-3334
dc.identifier.pmid2826035
dc.identifier.doi10.1093/carcin/9.1.81
dc.identifier.urihttp://hdl.handle.net/10541/115256
dc.description.abstractO6-alkylguanine-DNA-alkyltransferase (ATase)-deficient murine haemopoietic stem cells were transfected, following electroporation, with a G418-selectable expression vector containing the protein coding region of the Escherichia coli ATase gene ada. Clones of cells that were resistant to G418 or the chloroethylating agent mitozolomide (Mz) were selected and most were shown to express very high levels of bacterial gene-encoded ATase. In comparison with control cells that were transfected with the parent vector, the ATase-expressing clones were considerably more resistant to the toxic effects of the methylating agents N-methyl-N-nitrosourea and methylmethanesulphonate or the chloroethylating agents Mz or taurine chloroethylnitrosourea, but unchanged in their susceptibility to the bis-chloroethylating agent nitrogen mustard. Thus alkylation damage in DNA that can be repaired by the E. coli ATase constitutes the principal lethal lesion produced by alkylating agents in murine haemopoietic stem cells and the ATase deficiency in these cells can be complemented by electroporation-mediated gene transfection.
dc.language.isoenen
dc.subjectHaematopoietic Stem Cellsen
dc.subject.meshAlkyl and Aryl Transferases
dc.subject.meshAlkylating Agents
dc.subject.meshAnimals
dc.subject.meshDNA Restriction Enzymes
dc.subject.meshDeoxyribonuclease BamHI
dc.subject.meshDeoxyribonuclease EcoRI
dc.subject.meshDrug Resistance
dc.subject.meshEscherichia coli
dc.subject.meshFluorometry
dc.subject.meshHematopoietic Stem Cells
dc.subject.meshMechlorethamine
dc.subject.meshMethyl Methanesulfonate
dc.subject.meshMethylnitrosourea
dc.subject.meshMice
dc.subject.meshNitrogen Mustard Compounds
dc.subject.meshNitrosourea Compounds
dc.subject.meshPlasmids
dc.subject.meshTaurine
dc.subject.meshTransfection
dc.subject.meshTransferases
dc.titleTransfection of murine multi-potent haemopoietic stem cells with an E. coli DNA alkyltransferase gene confers resistance to the toxic effects of alkylating agents.en
dc.typeArticleen
dc.contributor.departmentDepartment of Experimental Haematology, Paterson Institute for Cancer Research, Christie Hospital, Manchester, UK.en
dc.identifier.journalCarcinogenesisen
html.description.abstractO6-alkylguanine-DNA-alkyltransferase (ATase)-deficient murine haemopoietic stem cells were transfected, following electroporation, with a G418-selectable expression vector containing the protein coding region of the Escherichia coli ATase gene ada. Clones of cells that were resistant to G418 or the chloroethylating agent mitozolomide (Mz) were selected and most were shown to express very high levels of bacterial gene-encoded ATase. In comparison with control cells that were transfected with the parent vector, the ATase-expressing clones were considerably more resistant to the toxic effects of the methylating agents N-methyl-N-nitrosourea and methylmethanesulphonate or the chloroethylating agents Mz or taurine chloroethylnitrosourea, but unchanged in their susceptibility to the bis-chloroethylating agent nitrogen mustard. Thus alkylation damage in DNA that can be repaired by the E. coli ATase constitutes the principal lethal lesion produced by alkylating agents in murine haemopoietic stem cells and the ATase deficiency in these cells can be complemented by electroporation-mediated gene transfection.


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