Methylmethane-sulphonate and X-ray-induced mutations in the Chinese hamster hprt gene: mRNA phenotyping using polymerase chain reactions.

2.50
Hdl Handle:
http://hdl.handle.net/10541/109874
Title:
Methylmethane-sulphonate and X-ray-induced mutations in the Chinese hamster hprt gene: mRNA phenotyping using polymerase chain reactions.
Authors:
Chaudhry, M Ahmad; Fox, Margaret
Abstract:
Alterations in the hprt gene of Chinese hamster cells were determined in 71 spontaneous, methylmethane sulphonate (MMS)- and X-ray-induced mutants, using the Southern blot hybridization technique. Among 41 MMS-induced mutants, deletions eliminating the whole gene were observed in 17 cases (41%). Analysis of 20 X-ray-induced mutants revealed the presence of similar deletions in nine of them (45%). No evidence of deletion was found in 10 spontaneous mutants. To investigate the possibility of small deletions, 18 MMS-induced mutants were studied with probes derived from exons 3 and 9 but no evidence of specific deletion of these two exons was found. The polymerase chain reaction (PCR) was used to phenotype hprt transcripts in 48 MMS, X-ray and spontaneous Chinese hamster mutants by amplifying the coding region of their cDNA. Among 22 MMS-induced mutants the message was present in 16 instances; 11 had normal levels while three had much lower levels of transcription. A further two mutants had mRNA of reduced size as revealed by the use of primers for PCR 3' to those routinely used. An analysis of 20 X-ray-induced mutants showed the presence of hprt mRNA in 11 of them with five having low levels of transcription. Among six spontaneous mutants, four were negative for mRNA on standard Northern blots and in one the message was only detected after PCR amplification. Direct DNA sequencing of 10 mutants revealed the presence of base substitutions in five of them while a 7 bp deletion was found in another. No mutations were found in another four mutants, suggesting the presence of mutation outside the coding region.
Affiliation:
CRC Department of Biochemical Genetics, Paterson Institute for Cancer Research, Christie Hospital and Holt Radium Institute, Manchester, UK.
Citation:
Methylmethane-sulphonate and X-ray-induced mutations in the Chinese hamster hprt gene: mRNA phenotyping using polymerase chain reactions. 1990, 5 (5):497-504 Mutagenesis
Journal:
Mutagenesis
Issue Date:
Sep-1990
URI:
http://hdl.handle.net/10541/109874
DOI:
10.1093/mutage/5.5.497
PubMed ID:
2263207
Type:
Article
Language:
en
ISSN:
0267-8357
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorChaudhry, M Ahmaden
dc.contributor.authorFox, Margareten
dc.date.accessioned2010-08-18T13:44:41Z-
dc.date.available2010-08-18T13:44:41Z-
dc.date.issued1990-09-
dc.identifier.citationMethylmethane-sulphonate and X-ray-induced mutations in the Chinese hamster hprt gene: mRNA phenotyping using polymerase chain reactions. 1990, 5 (5):497-504 Mutagenesisen
dc.identifier.issn0267-8357-
dc.identifier.pmid2263207-
dc.identifier.doi10.1093/mutage/5.5.497-
dc.identifier.urihttp://hdl.handle.net/10541/109874-
dc.description.abstractAlterations in the hprt gene of Chinese hamster cells were determined in 71 spontaneous, methylmethane sulphonate (MMS)- and X-ray-induced mutants, using the Southern blot hybridization technique. Among 41 MMS-induced mutants, deletions eliminating the whole gene were observed in 17 cases (41%). Analysis of 20 X-ray-induced mutants revealed the presence of similar deletions in nine of them (45%). No evidence of deletion was found in 10 spontaneous mutants. To investigate the possibility of small deletions, 18 MMS-induced mutants were studied with probes derived from exons 3 and 9 but no evidence of specific deletion of these two exons was found. The polymerase chain reaction (PCR) was used to phenotype hprt transcripts in 48 MMS, X-ray and spontaneous Chinese hamster mutants by amplifying the coding region of their cDNA. Among 22 MMS-induced mutants the message was present in 16 instances; 11 had normal levels while three had much lower levels of transcription. A further two mutants had mRNA of reduced size as revealed by the use of primers for PCR 3' to those routinely used. An analysis of 20 X-ray-induced mutants showed the presence of hprt mRNA in 11 of them with five having low levels of transcription. Among six spontaneous mutants, four were negative for mRNA on standard Northern blots and in one the message was only detected after PCR amplification. Direct DNA sequencing of 10 mutants revealed the presence of base substitutions in five of them while a 7 bp deletion was found in another. No mutations were found in another four mutants, suggesting the presence of mutation outside the coding region.en
dc.language.isoenen
dc.subject.meshAnimals-
dc.subject.meshBase Sequence-
dc.subject.meshBlotting, Northern-
dc.subject.meshBlotting, Southern-
dc.subject.meshCell Line-
dc.subject.meshExons-
dc.subject.meshGenes-
dc.subject.meshHypoxanthine Phosphoribosyltransferase-
dc.subject.meshMethyl Methanesulfonate-
dc.subject.meshMolecular Sequence Data-
dc.subject.meshMutation-
dc.subject.meshPhenotype-
dc.subject.meshPolymerase Chain Reaction-
dc.subject.meshRNA, Messenger-
dc.titleMethylmethane-sulphonate and X-ray-induced mutations in the Chinese hamster hprt gene: mRNA phenotyping using polymerase chain reactions.en
dc.typeArticleen
dc.contributor.departmentCRC Department of Biochemical Genetics, Paterson Institute for Cancer Research, Christie Hospital and Holt Radium Institute, Manchester, UK.en
dc.identifier.journalMutagenesisen
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