DNA sequence analysis of revertants of the hisD3052 allele of Salmonella typhimurium TA98 using the polymerase chain reaction and direct sequencing: Application to 1-nitropyrene-induced revertants
| Autor: | David M. DeMarini, Douglas A. Bell, Jessie G. Levine |
|---|---|
| Rok vydání: | 1991 |
| Předmět: |
Salmonella typhimurium
Salmonella Sequence analysis Molecular Sequence Data Biology Toxicology medicine.disease_cause Polymerase Chain Reaction law.invention symbols.namesake chemistry.chemical_compound law Genetics medicine Gene Polymerase chain reaction Electrophoresis Agar Gel Sanger sequencing Mutation Pyrenes Base Sequence Mutagenicity Tests Nucleic acid sequence DNA Molecular biology chemistry Genes Bacterial symbols Chromosome Deletion |
| Zdroj: | Mutation Research/Environmental Mutagenesis and Related Subjects. 252:35-44 |
| ISSN: | 0165-1161 |
| DOI: | 10.1016/0165-1161(91)90249-8 |
| Popis: | We have used the polymerase chain reaction (PCR) to speed the DNA sequence analysis of revertants of Salmonella typhimurium TA98. Briefly, a crude DNA extract from a single colony was prepared and used in an asymmetric PCR to amplify a 328-bp fragment containing the hisD3052 mutation approximately in the center. Following ultrafiltration, the ssDNA was sequenced using an end-labeled probe and dideoxy sequencing. The most frequent mutation among the revertants was a -2 deletion of GC or CG within the sequence CGCGCGCG, which is upstream of the hisD3052 mutation. This deletion occurred in 38% (6/16) of the spontaneous (-S9) revertants and in 94% (15/16) of a set of 1-nitropyrene-induced revertants. Other mutations, mostly deletions but also some complex mutations (i.e., single mutational events involving a combination of insertions, deletions, and substitutions), occurred within quasipalindromic regions of DNA. Possible mutational mechanisms are discussed, and the results with 1-NP are compared to those obtained in other systems. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|