Colony probing as an alternative to standard sequencing as a means of direct analysis of chromosomal DNA to determine the spectrum of single-base changes in regions of known sequence.
We demonstrate here the means to directly analyze bacterial chromosomal DNA for all classes of single-base mutations in a specific codon in any region of known sequence through the use of DNA probing as a powerful substitute for standard sequencing techniques. With this method, chromosomal DNA from hundreds of mutants can be examined for single-base changes without any DNA cloning. This method can conveniently provide a large data base for the assessment of all classes of single-base mutations occurring spontaneously or induced by a known or suspected mutagen. The method is demonstrated for the analysis of histidine-independent (His+) revertants of hisG46, a missense mutation of Salmonella typhimurium that can revert in six or seven ways.
J K
JK
Miller
W M
WM
Barnes
D001374:Azacitidine / Q000494:pharmacology; D001431:Bacteriological Techniques; D001483:Base Sequence; D002876:Chromosomes, Bacterial / Q000032:analysis; D003062:Codon; D004269:DNA, Bacterial / Q000032:analysis*; D005798:Genes, Bacterial; D005819:Genetic Markers; D005821:Genetic Techniques; D009154:Mutation; D009693:Nucleic Acid Hybridization; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis / Q000235:genetics; D012486:Salmonella typhimurium / Q000187:drug effects / Q000235:genetics
D003062:Codon; D004269:DNA, Bacterial; D005819:Genetic Markers; D009838:Oligodeoxyribonucleotides; D001374:Azacitidine