A second homologous recombination was promoted
by growth on M9 agar plates containing sucrose, which
retards the growth of bacteria expressing SacB. Generation of the
deletion mutation was confirmed by polymerase chain reaction
(PCR) amplification using primers HrcC.for (5′-TGTATTTCT
ACCCGGGCAAGTCC-3′) and HrcC.rev (5′-TCCTCGATGTAG
AGACTCAGCTTG-3′)