Analysis of spontaneous and psoralen-induced Salmonella typhimurium hisG46 revertants by oligodeoxyribonucleotide colony hybridization: use of psoralens to cross-link probes to target sequences.
Identifieur interne : 002A27 ( PubMed/Curation ); précédent : 002A26; suivant : 002A28Analysis of spontaneous and psoralen-induced Salmonella typhimurium hisG46 revertants by oligodeoxyribonucleotide colony hybridization: use of psoralens to cross-link probes to target sequences.
Auteurs : T A Cebula [États-Unis] ; W H KochSource :
- Mutation research [ 0027-5107 ] ; 1990.
Descripteurs français
- KwdFr :
- ADN bactérien (effets des radiations), ADN bactérien (génétique), Analyse de mutations d'ADN, Données de séquences moléculaires, Furocoumarines (pharmacologie), Hybridation d'acides nucléiques, Méthodes, Oligodésoxyribonucléotides (génétique), Psoralène (pharmacologie), Rayons ultraviolets, Reproductibilité des résultats, Réactifs réticulants (pharmacologie), Salmonella typhimurium (génétique), Séquence nucléotidique.
- MESH :
- effets des radiations : ADN bactérien.
- génétique : ADN bactérien, Oligodésoxyribonucléotides, Salmonella typhimurium.
- pharmacologie : Furocoumarines, Psoralène, Réactifs réticulants.
- Analyse de mutations d'ADN, Données de séquences moléculaires, Hybridation d'acides nucléiques, Méthodes, Rayons ultraviolets, Reproductibilité des résultats, Séquence nucléotidique.
English descriptors
- KwdEn :
- Base Sequence, Cross-Linking Reagents (pharmacology), DNA Mutational Analysis, DNA, Bacterial (genetics), DNA, Bacterial (radiation effects), Ficusin (pharmacology), Furocoumarins (pharmacology), Methods, Molecular Sequence Data, Nucleic Acid Hybridization, Oligodeoxyribonucleotides (genetics), Reproducibility of Results, Salmonella typhimurium (genetics), Ultraviolet Rays.
- MESH :
- chemical , genetics : DNA, Bacterial, Oligodeoxyribonucleotides.
- chemical , pharmacology : Cross-Linking Reagents, Ficusin, Furocoumarins.
- chemical , radiation effects : DNA, Bacterial.
- genetics : Salmonella typhimurium.
- Base Sequence, DNA Mutational Analysis, Methods, Molecular Sequence Data, Nucleic Acid Hybridization, Reproducibility of Results, Ultraviolet Rays.
Abstract
An improved DNA colony-hybridization method for the rapid characterization of Salmonella typhimurium hisG46 revertants is described. Oligodeoxyribonucleotides (15-mers) complementary to each of 6 possible transition or transversion mutations and an extragenic suppressor mutation, underlying the His+ phenotype, were prepared. Optimal sequence discrimination was achieved by hybridizing 15-mers at the apparent dissociation temperature (Td) for 2 h with chromosomal DNA of revertant colonies affixed to Whatman 541 filters. Subsequent exposure of filters to UVA radiation (320-400 nm) in the presence of 4'-hydroxymethyl-4,5',8-trimethylpsoralen (HMT) resulted in cross-linking of perfectly matched probes and target DNA sequences while sequences containing a single base-pair mismatch could be discriminated with a brief denaturing wash. No false negative results were obtained with the new procedure. An analysis of 204 spontaneous and 174 PUVA-induced TA100 revertants is presented.
DOI: 10.1016/0027-5107(90)90010-2
PubMed: 2179713
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Hybridization</term><term>Reproducibility of Results</term><term>Ultraviolet Rays</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Analyse de mutations d'ADN</term><term>Données de séquences moléculaires</term><term>Hybridation d'acides nucléiques</term><term>Méthodes</term><term>Rayons ultraviolets</term><term>Reproductibilité des résultats</term><term>Séquence nucléotidique</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">An improved DNA colony-hybridization method for the rapid characterization of Salmonella typhimurium hisG46 revertants is described. Oligodeoxyribonucleotides (15-mers) complementary to each of 6 possible transition or transversion mutations and an extragenic suppressor mutation, underlying the His+ phenotype, were prepared. Optimal sequence discrimination was achieved by hybridizing 15-mers at the apparent dissociation temperature (Td) for 2 h with chromosomal DNA of revertant colonies affixed to Whatman 541 filters. Subsequent exposure of filters to UVA radiation (320-400 nm) in the presence of 4'-hydroxymethyl-4,5',8-trimethylpsoralen (HMT) resulted in cross-linking of perfectly matched probes and target DNA sequences while sequences containing a single base-pair mismatch could be discriminated with a brief denaturing wash. No false negative results were obtained with the new procedure. An analysis of 204 spontaneous and 174 PUVA-induced TA100 revertants is presented.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">2179713</PMID><DateCompleted><Year>1990</Year><Month>04</Month><Day>20</Day></DateCompleted><DateRevised><Year>2019</Year><Month>07</Month><Day>02</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0027-5107</ISSN><JournalIssue CitedMedium="Print"><Volume>229</Volume><Issue>1</Issue><PubDate><Year>1990</Year><Month>Mar</Month></PubDate></JournalIssue><Title>Mutation research</Title><ISOAbbreviation>Mutat. 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Subsequent exposure of filters to UVA radiation (320-400 nm) in the presence of 4'-hydroxymethyl-4,5',8-trimethylpsoralen (HMT) resulted in cross-linking of perfectly matched probes and target DNA sequences while sequences containing a single base-pair mismatch could be discriminated with a brief denaturing wash. No false negative results were obtained with the new procedure. 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