Second-site reversion of a structural defect in bacteriophage T4 lysozyme.
A plasmid-borne gene encoding bacteriophage T4 lysozyme with a structural mutation, Tyr161-Ala, was mutagenized by by the use of polymerase chain reaction. The mutagenized gene was inserted into a specialized bacteriophage lambda cloning vector that must acquire a functional lysozyme gene in order to form plaques. Functional variants of the mutant lysozyme were selected. Three compensatory second-site revertants were obtained: Thr152-Met, Lys43-Ile, and Thr151-Ala. The effects of these mutations are interpreted in light of previous structural and genetic studies of T4 lysozyme.
Bouvier
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Department of Molecular Genetics and Microbiology, University of Massachusetts Medical Center, Worcester 01655, USA.
Poteete
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AR
D017122:Bacteriophage T4 / Q000201:enzymology*; D010582:Bacteriophage lambda / Q000235:genetics; D001483:Base Sequence; D005822:Genetic Vectors; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009113:Muramidase / Q000737:chemistry / Q000235:genetics*; D016296:Mutagenesis; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D017510:Protein Folding; D012641:Selection, Genetic; D017422:Sequence Analysis, DNA
D009113:Muramidase