Main Title |
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. |
Author |
Bell, D. A. ;
Levine, J. G. ;
DeMarini., D. M. ;
|
CORP Author |
Health Effects Research Lab., Research Triangle Park, NC. Genetic Toxicology Div. |
Publisher |
c1991 |
Year Published |
1991 |
Report Number |
EPA/600/J-91/098; |
Stock Number |
PB91-200329 |
Additional Subjects |
Bacterial DNA ;
Salmonella typhimurium ;
Mutation ;
Polymerase chain reaction ;
Alleles ;
Base sequence ;
Metabolic activation ;
Single-stranded DNA ;
Nucleic acid conformation ;
Agar gel electrophoresis ;
Reprints ;
1-Nitropyrene
|
Holdings |
Library |
Call Number |
Additional Info |
Location |
Last Modified |
Checkout Status |
NTIS |
PB91-200329 |
Some EPA libraries have a fiche copy filed under the call number shown. |
|
07/26/2022 |
|
Collation |
12p |
Abstract |
The study used the polymerase chain reaction (PCR) to speed the processing of revertants of Salmonella typhimurium TA98 for DNA sequence analysis. Briefly, a crude DNA extract from a single colony was prepared and used in an asymmetric PCR to amplify a 228-bp fragment containing the hisD3052 mutation approximately in the center. Following ultra-filtration, 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. The deletion occurred in 38% (6/16) of the spontaneous (-S9) revertants and in 94% (15/16) of a set of 1-nitropyrene-induced revertants. Misalignment of complementary DNA strands within this repeat may account for this mutation, although the possible formation of Z-DNA within this region may also play a role. Other mutations, mostly deletions but also some complex mutations (insertions/deletions/substitutions), occurred within quasi-palindromic regions of DNA. The potential DNA secondary structures within such regions may mediate the templated production of some of these mutations. |
Supplementary Notes |
Pub. in Mutation Research, v252 n1 p35-44 Feb 91. |
NTIS Title Notes |
Journal article. |
Title Annotations |
Reprint: 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. |
Category Codes |
57K; 57F |
NTIS Prices |
PC A03/MF A01 |
Primary Description |
600/10 |
Document Type |
NT |
Cataloging Source |
NTIS/MT |
Control Number |
122131013 |
Origin |
NTIS |
Type |
CAT |