Isolation of bacteriophage T4 DNA polymerase mutator mutants.

Abstract:

More than 20 new bacteriophage T4 DNA polymerase mutants have been isolated by a procedure designed to select mutants with high spontaneous mutation rates. Some of the mutants produce the highest mutation frequencies that have been observed in T4 thus far. The design of the selection procedure allows for the isolation of mutator mutants that preferentially induce certain types of replication errors, and some of the mutator mutants have mutational specificities different from wild-type. The new mutants are clustered at just two sites in the DNA polymerase gene, and this result confirms an earlier observation.

Polymerases:

Topics:

Mutational Analysis, Structure and Structure/Function, Fidelity, Nucleotide Incorporation, Exonuclease Activity, Methods

One line summary:

T4 DNA mutator mutant polymerases discovered by described method are clustered at just two sites in the gene.

Status:

new topics/pols set partial results complete validated

Results:

Polymerase Reference Property Result Context
T4 D131G Reha-Krantz LJ1986 Nucleotide Substitution Rate 2728 Mutation frequency (relative to WT) Technique: ac forward mutational assay
T4 G82D Reha-Krantz LJ1986 Nucleotide Substitution Rate 300 Mutation frequency (relative to WT) Technique: ac forward mutational assay
T4 G82D Reha-Krantz LJ1986 Nucleotide Substitution Rate 92 Mutation frequency (relative to WT) Technique: T4 rII P7oc reversion assay (AT->GC (P7oc -> P7am))
T4 G82D Reha-Krantz LJ1986 Nucleotide Substitution Rate 100 Mutation frequency (relative to WT) Technique: T4 rII P7oc reversion assay (AT->TA(CG) (P7oc->rII+))
T4 G82D Reha-Krantz LJ1986 Nucleotide Substitution Rate 112 Mutation frequency (relative to WT) Technique: T4 rII P7am reversion assay (GC->TA(CG) (P7am->rII+))
T4 L340P Reha-Krantz LJ1986 Nucleotide Substitution Rate 380 Mutation frequency (relative to WT) Technique: T4 rII P7oc reversion assay (AT->GC (P7oc->P7am))
T4 L340P Reha-Krantz LJ1986 Nucleotide Substitution Rate 320 Mutation frequency (relative to WT) Technique: T4 rII P7oc reversion assay (AT->TA(CG) (P7oc->rII+))
T4 L340P Reha-Krantz LJ1986 Nucleotide Substitution Rate 1125 Mutation frequency (relative to WT) Technique: T4 rII P7am reversion assay (GC->TA(CG) (P7am->rII+))
T4 A89TD363N Reha-Krantz LJ1986 Nucleotide Substitution Rate 90 Mutation frequency (relative to WT) Technique: T4 rII P7oc reversion assay (AT->GC (P7oc->P7am))
T4 A89TD363N Reha-Krantz LJ1986 Nucleotide Substitution Rate 570 Mutation frequency (relative to WT) Technique: T4 rII P7oc reversion assay (AT->TA(CG) (P7oc->rII+))
T4 A89TD363N Reha-Krantz LJ1986 Nucleotide Substitution Rate 1900 Mutation frequency (relative to WT) Technique: T4 rII P7am reversion assay (GC->TA(CG) (P7am->rII+))
T4 Y317C Reha-Krantz LJ1986 Nucleotide Substitution Rate 800 Mutation frequency (relative to WT) Technique: T4 rII P7oc reversion assay (AT->GC (P7oc->P7am))
T4 Y317C Reha-Krantz LJ1986 Nucleotide Substitution Rate 1720 Mutation frequency (relative to WT) Technique: T4 rII P7oc reversion assay (AT->TA(CG) (P7oc->rII+))
T4 Y317C Reha-Krantz LJ1986 Nucleotide Substitution Rate 2110 Mutation frequency (relative to WT) Technique: T4 rII P7am reversion assay (GC->TA(CG) (P7am->rII+))
T4 Q478RA488V Reha-Krantz LJ1986 Nucleotide Substitution Rate 266 Mutation frequency (relative to WT) Technique: T4 rII P7oc reversion assay (AT->GC (P7oc->P7am))
T4 Q478RA488V Reha-Krantz LJ1986 Nucleotide Substitution Rate 200 Mutation frequency (relative to WT) Technique: T4 rII P7oc reversion assay (AT->TA(CG) (P7oc->rII+))
T4 Q478RA488V Reha-Krantz LJ1986 Nucleotide Substitution Rate 188 Mutation frequency (relative to WT) Technique: T4 rII P7am reversion assay (GC->TA(CG) (P7am->rII+))

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