Analysis of Salmonella typhimurium hisD3052 revertants: the use of oligodeoxyribonucleotide colony hybridization, PCR, and direct sequencing in mutational analysis

E Kupchella; T A Cebula

doi:10.1002/em.2850180404

Analysis of Salmonella typhimurium hisD3052 revertants: the use of oligodeoxyribonucleotide colony hybridization, PCR, and direct sequencing in mutational analysis

Environ Mol Mutagen. 1991;18(4):224-30. doi: 10.1002/em.2850180404.

Authors

E Kupchella¹, T A Cebula

Affiliation

¹ Molecular Biology Branch, Food and Drug Administration, Washington, D.C. 20204.

PMID: 1748083
DOI: 10.1002/em.2850180404

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 oligodeoxyribonucleotide 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.

MeSH terms

Base Sequence
DNA Mutational Analysis*
Histidine
Molecular Sequence Data
Mutagenicity Tests*
Nucleic Acid Hybridization
Oligonucleotide Probes / chemistry
Polymerase Chain Reaction
Salmonella typhimurium / genetics*

Substances

Oligonucleotide Probes
Histidine