Isolation and cDNA cloning of a rat O6-alkylguanine-DNA-alkyltransferase gene, molecular analysis of expression in rat liver.

2.50
Hdl Handle:
http://hdl.handle.net/10541/104602
Title:
Isolation and cDNA cloning of a rat O6-alkylguanine-DNA-alkyltransferase gene, molecular analysis of expression in rat liver.
Authors:
Potter, P M; Rafferty, Joseph A; Cawkwell, L; Wilkinson, M C; Cooper, Donald P; O'Connor, Peter J; Margison, Geoffrey P
Abstract:
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.
Affiliation:
CRC Department of Chemical Carcinogenesis, Paterson Institute for Cancer Research, Manchester, UK.
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 Carcinogenesis
Journal:
Carcinogenesis
Issue Date:
Apr-1991
URI:
http://hdl.handle.net/10541/104602
DOI:
10.1093/carcin/12.4.727
PubMed ID:
2013136
Type:
Article
Language:
en
ISSN:
0143-3334
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorPotter, P Men
dc.contributor.authorRafferty, Joseph Aen
dc.contributor.authorCawkwell, Len
dc.contributor.authorWilkinson, M Cen
dc.contributor.authorCooper, Donald Pen
dc.contributor.authorO'Connor, Peter Jen
dc.contributor.authorMargison, Geoffrey Pen
dc.date.accessioned2010-06-10T09:02:56Z-
dc.date.available2010-06-10T09:02:56Z-
dc.date.issued1991-04-
dc.identifier.citationIsolation and cDNA cloning of a rat O6-alkylguanine-DNA-alkyltransferase gene, molecular analysis of expression in rat liver. 1991, 12 (4):727-33 Carcinogenesisen
dc.identifier.issn0143-3334-
dc.identifier.pmid2013136-
dc.identifier.doi10.1093/carcin/12.4.727-
dc.identifier.urihttp://hdl.handle.net/10541/104602-
dc.description.abstractA 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.en
dc.language.isoenen
dc.subject.mesh2-Acetylaminofluorene-
dc.subject.meshAmino Acid Sequence-
dc.subject.meshAnimals-
dc.subject.meshBase Sequence-
dc.subject.meshBlotting, Southern-
dc.subject.meshCloning, Molecular-
dc.subject.meshDNA-
dc.subject.meshEscherichia coli-
dc.subject.meshGene Expression-
dc.subject.meshHumans-
dc.subject.meshLiver-
dc.subject.meshMethyltransferases-
dc.subject.meshMolecular Sequence Data-
dc.subject.meshO(6)-Methylguanine-DNA Methyltransferase-
dc.subject.meshRNA, Messenger-
dc.subject.meshRats-
dc.subject.meshRats, Inbred Strains-
dc.subject.meshSequence Homology, Nucleic Acid-
dc.subject.meshUp-Regulation-
dc.titleIsolation and cDNA cloning of a rat O6-alkylguanine-DNA-alkyltransferase gene, molecular analysis of expression in rat liver.en
dc.typeArticleen
dc.contributor.departmentCRC Department of Chemical Carcinogenesis, Paterson Institute for Cancer Research, Manchester, UK.en
dc.identifier.journalCarcinogenesisen

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