Abstract
We developed a novel negative selection system for actinobacteria based on cytosine deaminase (CodA). We constructed vectors that include a synthetic gene encoding the CodA protein from Escherichia coli optimized for expression in Streptomyces species. Gene disruption and the introduction of an unmarked in-frame deletion were successfully achieved with these vectors.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Actinobacteria / genetics*
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Anti-Bacterial Agents / pharmacology
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Cytosine Deaminase / genetics
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Cytosine Deaminase / metabolism*
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Escherichia coli Proteins / genetics
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Escherichia coli Proteins / metabolism*
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Flucytosine / metabolism
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Flucytosine / pharmacology
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Fluorouracil / metabolism
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Fluorouracil / pharmacology
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Genetic Vectors
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Microbial Sensitivity Tests
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Mutagenesis, Insertional / methods*
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Selection, Genetic*
Substances
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Anti-Bacterial Agents
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Escherichia coli Proteins
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Flucytosine
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Cytosine Deaminase
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codA protein, E coli
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Fluorouracil