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 |
a
Underlining indicates restriction endonuclease site.