Isolation and cDNA cloning of a rat O6-alkylguanine-DNA-alkyltransferase gene, molecular analysis of expression in rat liver.
Authors
Potter, P MRafferty, Joseph A
Cawkwell, L
Wilkinson, M C
Cooper, Donald P
O'Connor, Peter J
Margison, Geoffrey P
Affiliation
CRC Department of Chemical Carcinogenesis, Paterson Institute for Cancer Research, Manchester, UK.Issue Date
1991-04
Metadata
Show full item recordAbstract
A rat O6-alkylguanine-DNA-alkyltransferase (ATase) cDNA has been isolated from a rat liver cDNA library by hybridization with the human homologue. The candidate 806 bp cDNA was sequenced and shown to contain a 630 bp open reading frame that could encode a protein of 22.2 kd. Fluorography of labelled ATase indicates a 24 kd protein which is consistent with several previous reports. The derived amino acid sequence demonstrated 81% similarity with the human ATase and 94% identity over a 67 residue region encompassing the putative alkyl acceptor site. Peptide sequences derived from cleaved homogeneous rat ATase have been located in the predicted protein providing additional confirmation of the identity of the cDNA. A 1.05 kb mRNA has been detected in rat liver by Northern analysis; treatment of adult rats with 2-acetylaminofluorene causes an approximately 10-fold induction of this message in liver. Following site directed mutagenesis of the 806 bp cDNA, the 630 bp protein coding sequence has been ligated into an Escherichia coli expression vector to achieve ATase levels of greater than 3% of total protein in bacterial extracts.Citation
Isolation and cDNA cloning of a rat O6-alkylguanine-DNA-alkyltransferase gene, molecular analysis of expression in rat liver. 1991, 12 (4):727-33 CarcinogenesisJournal
CarcinogenesisDOI
10.1093/carcin/12.4.727PubMed ID
2013136Type
ArticleLanguage
enISSN
0143-3334ae974a485f413a2113503eed53cd6c53
10.1093/carcin/12.4.727
Scopus Count
Collections
Related articles
- Characterisation and nucleotide sequence of ogt, the O6-alkylguanine-DNA-alkyltransferase gene of E. coli.
- Authors: Potter PM, Wilkinson MC, Fitton J, Carr FJ, Brennand J, Cooper DP, Margison GP
- Issue date: 1987 Nov 25
- cDNA cloning of the rat O6-methylguanine-DNA-methyltransferase.
- Authors: Rahden-Staron I, Laval F
- Issue date: 1991 Jun 14
- Isolation and partial characterization of murine O6-alkylguanine-DNA-alkyltransferase: comparative sequence and structural properties.
- Authors: Santibanez-Koref M, Elder RH, Fan CY, Cawkwell L, McKie JH, Douglas KT, Margison GP, Rafferty JA
- Issue date: 1992
- Expression of a human O6-alkylguanine-DNA-alkyltransferase cDNA in human cells and transgenic mice.
- Authors: Fan CY, Potter PM, Rafferty J, Watson AJ, Cawkwell L, Searle PF, O'Connor PJ, Margison GP
- Issue date: 1990 Oct 11
- Induction of O6-alkylguanine-DNA-alkyltransferase in the hepatocytes of rats following treatment with 2-acetylaminofluorene.
- Authors: Chinnasamy N, Rafferty JA, Margison GP, O'Connor PJ, Elder RH
- Issue date: 1997 Apr