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dc.contributor.authorFan, Chun-Yangen
dc.contributor.authorPotter, P Men
dc.contributor.authorRafferty, Joseph Aen
dc.contributor.authorWatson, Amanda Jen
dc.contributor.authorCawkwell, Len
dc.contributor.authorSearle, P Fen
dc.contributor.authorO'Connor, Peter Jen
dc.contributor.authorMargison, Geoffrey Pen
dc.date.accessioned2010-08-18T08:45:14Z
dc.date.available2010-08-18T08:45:14Z
dc.date.issued1990-10-11
dc.identifier.citationExpression of a human O6-alkylguanine-DNA-alkyltransferase cDNA in human cells and transgenic mice. 1990, 18 (19):5723-7 Nucleic Acids Res.en
dc.identifier.issn0305-1048
dc.identifier.pmid2216765
dc.identifier.doi10.1093/nar/18.19.5723
dc.identifier.urihttp://hdl.handle.net/10541/109801
dc.description.abstractA truncated human O6-alkylguanine-DNA-alkyltransferase (ATase) cDNA was ligated into an expression vector under the control of the mouse metallothionein-1 gene promotor and upstream of part of the human growth hormone gene to provide splice and polyadenylation signals. Transfection of this construct into human cells resulted in very high levels of ATase expression (more than 300 fmoles/mg protein versus less than 2 fm/mg protein in parent vector transfected control cells). Microinjection of a 4.2 kb fragment of this vector into B6D2F2 mouse embryos and implantation of survivors into pseudopregnant females has so far generated 35 offspring. Southern analysis of tail tip DNA has shown that 11 of the offspring are transgenic for the human ATase gene, between 1 and at least 30 copies of the gene being detected. Human ATase transcripts were detected in total RNA extracted from liver obtained from two male transgenic mice by partial hepatectomy. Cell free extracts of liver samples from five transgenic mice showed up to 4 times higher ATase levels than control livers.
dc.language.isoenen
dc.subject.meshAnimals
dc.subject.meshBase Sequence
dc.subject.meshBlotting, Southern
dc.subject.meshCell Line
dc.subject.meshCloning, Molecular
dc.subject.meshDNA
dc.subject.meshFemale
dc.subject.meshGene Expression Regulation, Enzymologic
dc.subject.meshGenetic Vectors
dc.subject.meshHepatectomy
dc.subject.meshHumans
dc.subject.meshLiver
dc.subject.meshMale
dc.subject.meshMethyltransferases
dc.subject.meshMice
dc.subject.meshMice, Transgenic
dc.subject.meshMicroinjections
dc.subject.meshMolecular Sequence Data
dc.subject.meshO(6)-Methylguanine-DNA Methyltransferase
dc.subject.meshTransfection
dc.titleExpression of a human O6-alkylguanine-DNA-alkyltransferase cDNA in human cells and transgenic mice.en
dc.typeArticleen
dc.contributor.departmentDepartment of Chemical Carcinogenesis, Paterson Institute for Cancer Research, Manchester, UK.en
dc.identifier.journalNucleic Acids Researchen
html.description.abstractA truncated human O6-alkylguanine-DNA-alkyltransferase (ATase) cDNA was ligated into an expression vector under the control of the mouse metallothionein-1 gene promotor and upstream of part of the human growth hormone gene to provide splice and polyadenylation signals. Transfection of this construct into human cells resulted in very high levels of ATase expression (more than 300 fmoles/mg protein versus less than 2 fm/mg protein in parent vector transfected control cells). Microinjection of a 4.2 kb fragment of this vector into B6D2F2 mouse embryos and implantation of survivors into pseudopregnant females has so far generated 35 offspring. Southern analysis of tail tip DNA has shown that 11 of the offspring are transgenic for the human ATase gene, between 1 and at least 30 copies of the gene being detected. Human ATase transcripts were detected in total RNA extracted from liver obtained from two male transgenic mice by partial hepatectomy. Cell free extracts of liver samples from five transgenic mice showed up to 4 times higher ATase levels than control livers.


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