Expression of a human O6-alkylguanine-DNA-alkyltransferase cDNA in human cells and transgenic mice.
Potter, P M
Rafferty, Joseph A
Watson, Amanda J
Searle, P F
O'Connor, Peter J
Margison, Geoffrey P
AffiliationDepartment of Chemical Carcinogenesis, Paterson Institute for Cancer Research, Manchester, UK.
MetadataShow full item record
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.
CitationExpression of a human O6-alkylguanine-DNA-alkyltransferase cDNA in human cells and transgenic mice. 1990, 18 (19):5723-7 Nucleic Acids Res.
JournalNucleic Acids Research
- Tissue-specific expression and induction of human O6-alkylguanine-DNA alkyltransferase in transgenic mice.
- Authors: Rafferty JA, Fan CY, Potter PM, Watson AJ, Cawkwell L, O'Connor PJ, Margison GP
- Issue date: 1992
- Isolation and cDNA cloning of a rat O6-alkylguanine-DNA-alkyltransferase gene, molecular analysis of expression in rat liver.
- Authors: Potter PM, Rafferty JA, Cawkwell L, Wilkinson MC, Cooper DP, O'Connor PJ, Margison GP
- Issue date: 1991 Apr
- High level, regulated expression of the chimeric P-enolpyruvate carboxykinase (GTP)-bacterial O6-alkylguanine-DNA alkyltransferase (ada) gene in transgenic mice.
- Authors: Lim IK, Dumenco LL, Yun J, Donovan C, Warman B, Gorodetzkaya N, Wagner TE, Clapp DW, Hanson RW, Gerson SL
- Issue date: 1990 Mar 15
- Cell type-specific expression of the O6-alkylguanine-DNA alkyltransferase gene in normal human liver tissues as revealed by in situ hybridization.
- Authors: Wani G, Wani AA, D'Ambrosio SM
- Issue date: 1993 Apr
- Expression in mammalian cells of the Escherichia coli O6 alkylguanine-DNA-alkyltransferase gene ogt reduces the toxicity of alkylnitrosoureas.
- Authors: Harris LC, Margison GP
- Issue date: 1993 Jun