One hrcC deletion mutant, designated
AAC00-1ΔhrcC, was suitable for our studies. The hrcC deletion
mutant was complemented by subcloning the 3.6-kb BamHI fragment
containing the hrcC gene into the cosmid pUFR043 to produce
pUFR034-hrcC. pUFR043-hrcC was subsequently transformed
into E. coli S17-1 (λpir) for use in triparental mating.
pUFR043-hrcC was then transformed into AAC00-1ΔhrcC to create
AAC00-1ΔhrcCcomp (hrcC mutation complement).