TABLE 2.

Primers used in this study

Primer	Sequence (5′-3′)a	Purpose
vacB-N	ATGATCCTCGACCTCATCAAGGGCCATGAA	PCR amplification of a portion (2.0 kb) of the vacB gene
vacB-C	TCACGAAGTCGATCTTGCGATCGTCCAGAT
vacB-N1	TGCTGGTCTCGGCGAACAGGTGCCG	DNA sequencing of the PCR product to obtain middle part of the vacB gene
vacB-C1	CACCAGACCGTCGATGTGGATCTCG
vacB-N/TGA	CTCACCAACGACTACTACCAGTTCGACCCG	DNA sequencing of the gDNA to obtain the ORF of the vacB gene
vacB-C/ATG	CAGTTGACCGTCGCGCTCCATGGCGCGCAG
vacB-N/up	GTCTCATCATGACTGCAAACTGTGCATAGTATGAC	Cloning of the vacB gene into pCR2.1 vector
vacB-C/down	GGGGCCATCAGCCCCTTACGACCTCAGACCGGGTC
vacB-N/g	GAACGCCTGCTTGAGATAGGCGGCGGATTGGGCGA	PCR verification of the vacB mutant
vacB-C/g	AGTAACAGGGTTCGGATTTTTTTCGTTTTCAGGAC
vacB/HindIIIN	CCCAAGCTTGTCTCATCATGACTGCAAACTGTGCA	Cloning of the vacB gene into pBR322 vector
vacB/SalIC	ACGCGTCGACGGGGCCATCAGCCCCTTACGACCTC
vacB/AflIIIN	CCCACATGTCTCAAAAAGATCCTTTCCTCGAACGCGAGGC	Cloning of the vacB gene into pBAD/Thio-E vector
vacB/PmeIC	AGCTTTGTTTAAACTTACCCCTTGGCCTTGTCGCGCGCTT

^aUnderlining indicates restriction endonuclease site.