Some Strategies for Finding Mutants
"You get what you select for, but you don't know what you are selecting for until you get it." Tom Silhavy
Isolation of mutants in catabolic pathways
- Selection for resistance to a toxic substrate analog
- Selection for increased expression (e.g. faster growth on limiting substrate)
- Screen for failure to grow on a particular carbon or nitrogen source
Isolation of mutants in biosynthetic pathways
- Penicillin enrichment
- Screen for failure to grow without a particular nutritional supplement
Isolation of mutants in essential genes
- Screen for conditional lethal mutants
- Brute force screen for loss of an enzymatic activity
Isolation of mutants in defined loci or regions of the chromosome
- Localized mutagenesis
- Allele replacement
- Subtractive mutagenesis
- Select or screen for mutants in essential genes in diploids
Isolation of mutations that affect complex interactions
- Selection for suppressor mutations
- Screen for synthetic mutations
- Selection or screen for increased or decreased expression of reporter genes
- Beckwith, J. 1991. Strategies for finding mutants. Methods Enzymol. 204: 1-18.
- Shuman H., and T. Silhavy. 2003. The art and design of genetic screens: Escherichia coli.
Nat Rev Genet. 4: 419-431.
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Last modified July 2, 2004