Fig. 2.

Molecular cloning strategy for generating transformation substrates used to cis-complement mutated genes by genetic reconstitution. Small arrows represent primers with the relevant restriction enzyme sites (AatII and AgeI); the large black arrow represents the gene of interest to be complemented; the large dark gray arrow represents the gene conferring antibiotic resistance, which must be different than the antibiotic resistance gene used to mutate the gene of interest, although the promoter (P) driving expression can be the same; and the large light gray boxes represent the flanking upstream (up) and downstream (down) sequences for recombination.