Identification of the pheS5 mutation, which causes thermosensitivity of Escherichia coli mutant NP37.
The pheS5 mutation responsible for the thermosensitive phenylalanyl-tRNA synthetase of the classical Escherichia coli NP37 was cloned by a recombination event and identified by DNA sequence analysis. The mutation was subsequently verified by direct sequencing of amplified NP37 DNA generated by an asymmetric polymerase chain reaction. The resulting amino acid exchange, Gly-98 to Asp-98 in the phenylalanyl-tRNA synthetase alpha subunit, might cause subunit disaggregation due to electrostatic repulsion.
Kast
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Mikrobiologisches Institut, Eidgenössische Technische Hochschule, ETH-Zentrum, Zürich, Switzerland.
Keller
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Hennecke
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D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005816:Genetic Complementation Test; D016296:Mutagenesis; D009154:Mutation; D010652:Phenylalanine-tRNA Ligase / Q000235:genetics*; D016133:Polymerase Chain Reaction; D013696:Temperature
D010652:Phenylalanine-tRNA Ligase