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Analysis of Salmonella typhimurium hisD3052 revertants: The use of oligodeoxyribonucleotide colony hybridization, PCR, and direct sequencing in mutational analysis
Author(s) -
Kupchella Eugene,
Cebula Thomas A.
Publication year - 1991
Publication title -
environmental and molecular mutagenesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1
H-Index - 87
eISSN - 1098-2280
pISSN - 0893-6692
DOI - 10.1002/em.2850180404
Subject(s) - biology , polymerase chain reaction , microbiology and biotechnology , dna , genetics , dna sequencing , nucleic acid thermodynamics , salmonella , gene , base sequence , bacteria
Abstract A rapid method for determining the DNA sequences of Salmonella typhimurium hisD3052 revertants is presented. DNA colony hybridization was used to analyze revertants previously studied by Isono and Yourno [Proc Natl Acad Sci USA 71:1612–1617, 1974]. Synthetic oligodeoxy‐ribonucleotide probes (18‐mers) were able to distinguish sequences that differed by a single base pair. Mutant his sequences not identified by probing analysis were amplified using polymerase chain reaction (PCR) and directly sequenced. The combined use of DNA‐colony hybridization and direct sequencing offers a precise and rapid means for the molecular characterization of hisD3052 revertants.