Frameshift mutagenesis in bacteriophage T7.
We have undertaken an initial characterization of frameshift mutagenesis in bacteriophage T7 and have identified a subset of very low reversion frameshift mutations in the T7 ligase gene (gene 1.3). We used this information to construct bacteriophage T7 strains that contain one extra or one less base pair in gene 1.3 such that a frameshift event restores the reading frame of that gene. These events can be quantified and the frameshift mutation isolated within a localized region of the ligase gene. We have also identified a portion of the T7 ligase protein that will accept tracts of nonsense amino acids yet still give a ligase positive phenotype. This allows flexibility in the design of the target DNA sequence with which to study frameshift mutagenesis. These assays for frameshift mutagenesis performed in E. coli cells infected with the appropriate T7 strain, were used to measure the frequency of both plus and minus frameshifts in vivo.
Temple University School of Medicine, Department of Biochemistry and Molecular Biology, Philadelphia, PA 19140.
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JC
Pierce
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Masker
D000595:Amino Acid Sequence; D001482:Base Composition / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D011088:DNA Ligases / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D016368:Frameshift Mutation / Q000235:genetics*; D008969:Molecular Sequence Data; D016296:Mutagenesis / Q000235:genetics*; D015183:Restriction Mapping; D013604:T-Phages / Q000201:enzymology* / Q000235:genetics
D011088:DNA Ligases