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Table 1 List of mutagenesis and selection media for attempts to generate a null rpoN mutant.

From: Genome-wide analysis of the RpoN regulon in Geobacter sulfurreducens

Mutagenesis

Selection medium (electron donor/acceptor)

1. Deletion and replacement of the rpoN gene by double-crossover (Figure 1b)

Acetate/fumarate

 

Acetate/Fe(III) citrate

 

Acetate/Fe(III) citrate amended with glutamine

 

Acetate/Fe(III) citrate amended with glutamate

 

Acetate/Fe(III) citrate amended with both glutamine and glutamate

2. Deletion and replacement of the 5'-end of the rpoN gene by double-crossover (Figure 1b)

Acetate/fumarate

 

Acetate/Fe(III) citrate

3. Deletion and replacement of the 3'-end of the rpoN gene by double-crossover (Figure 1b)

Acetate/fumarate

 

Acetate/Fe(III) citrate

4. Deletion and replacement of the upstream intergenic region of the rpoN gene by double-crossover (Figure 1a)

Acetate/fumarate

5. Deletion and replacement of the downstream intergenic region of the rpoN gene by double-crossover (Figure 1a)

Acetate/fumarate

6. Integration of a linear DNA fragment on the chromosome, providing another copy of the rpoN gene on another location of the chromosome (Figure 1c)

Acetate/fumarate

 

Acetate/Fe(III) citrate

7. Integration of a linear DNA fragment on the chromosome, providing another copy of the rpoN gene in trans, which is either constitutively expressed by a lac promoter, or is IPTG inducible (Figure 1d)

Acetate/fumarate

 

Acetate/Fe(III) citrate

 

Acetate/Fe(III) citrate amended with both glutamine and glutamate