Isolation of bacteriophage T4 DNA polymerase mutator mutants.

Reha-Krantz LJ, Liesner EM, Parmaksizoglu S, Stocki S

Journal of molecular biology (1986), Volume 189, Page 261

PubMed entry

Abstract:

More than 20 new bacteriophage T4 DNA polymerase mutants have been ...

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:

T4 D131G,T4 Q478RA488V,T4 A89TD363N,T4,T4 G694S,T4 Y317C,T4 L340P,T4 S345P,T4 G82D

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:

Results:

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