Work

Escherichia coli thymidylate synthase: amino acid substitutions by suppression of amber nonsense mutations.

Year: 1990

Type: article

Abstract: By using site-directed oligonucleotide mutagenesis, amber nonsense stop codons (5'-TAG-3') have been introduced at 20 sites in the Escherichia coli thymidylate synthase gene. By transforming the thyA mutant plasmids into 13 strains, each of which harbor different amber suppressor tRNAs, we were able to generate over 245 amino acid substitutions in E. coli thymidylate synthase (EC 2.1.1.45). Growth characteristics of these mutants have been studied, yielding a body of information that includes some surprising results in light of the recently published crystal structure of the enzyme.

Source: Proceedings of the National Academy of Sciences

DOI: 10.1073/pnas.87.10.3957

Authors: M L Michaels, C W Kim, D A Matthews, J H Miller

Institution: University of California, Los Angeles

Language: English

Topic: Biochemical and Molecular Research

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: bronze

Funders: National Science Foundation, U.S. Public Health Service, National Institute of General Medical Sciences

Awards: Predoctoral Training Program in Genetic Mechanisms, Collaborative Research: Construction and Use of E. Coli Nonsense Suppressor Bank, G894887781