Expression of a human O6-alkylguanine-DNA-alkyltransferase cDNA in human cells and transgenic mice.

2.50
Hdl Handle:
http://hdl.handle.net/10541/109801
Title:
Expression of a human O6-alkylguanine-DNA-alkyltransferase cDNA in human cells and transgenic mice.
Authors:
Fan, Chun-Yang; Potter, P M; Rafferty, Joseph A; Watson, Amanda J; Cawkwell, L; Searle, P F; O'Connor, Peter J; Margison, Geoffrey P
Abstract:
A 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.
Affiliation:
Department of Chemical Carcinogenesis, Paterson Institute for Cancer Research, Manchester, UK.
Citation:
Expression of a human O6-alkylguanine-DNA-alkyltransferase cDNA in human cells and transgenic mice. 1990, 18 (19):5723-7 Nucleic Acids Res.
Journal:
Nucleic Acids Research
Issue Date:
11-Oct-1990
URI:
http://hdl.handle.net/10541/109801
DOI:
10.1093/nar/18.19.5723
PubMed ID:
2216765
Type:
Article
Language:
en
ISSN:
0305-1048
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
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.en
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

Related articles on PubMed

All Items in Christie are protected by copyright, with all rights reserved, unless otherwise indicated.