TABLE 2.
Oligonucleotides used in this study
| Name | Oligonucleotide Sequence (5′ → 3′)a,b | Use |
|---|---|---|
| For mutant construction | ||
| GBPA01-F | GAGATGCACATCAGCAACGCG | Deletion of the gbpA ORF |
| GBPA01-R | AAAGGATCCAGCGAACTTACACAGTGT | |
| GBPA02-F | GCTGGATCCTTTGTCTTCCCAGATG | Deletion of the gbpA ORF |
| GBPA02-R | CGCAACAACGGAATCAAACGC | |
| For mutant complementation | ||
| GBPA03-F | GGATCCGCCAAATAAAGTCAG | Amplification of the gbpA ORF |
| GBPA03-R | GGATCCTTACAGTTTGTCCCAC | |
| ISCR01-F | ATCCATGGCTATGAAACTGACATCTAAAGG | Amplification of the iscR ORF |
| ISCR01-R | ATTCTAGATTAAGAGCGGAAATTTACACCG | |
| CRP01-F | GAGATACCATGGTTCTAGGTAAACCTCA | Amplification of the crp ORF |
| CRP01-R | GTTAATTCTAGATTAACGAGTACCGTAAACAAC | |
| SMCR01-F | ATCCATGGACTCAATCGCAAAGAGAC | Amplification of the smcR ORF |
| SMCR02-R | ATTCTAGATTATTCGTGCTCGCGTTTATA | |
| For GbpA overexpression | ||
| GBPA04-F | CCATGGCTAAAAAACAACCGCAAAAAACC | Amplification of the gbpA ORF |
| GBPA04-R | CTCGAGCAGTTTGTCCCACGCCATT | |
| For promoter deletion analysis | ||
| GBPA003 | GAGCTCTAAGTGCTCAATGACATAGTAAAG | Deletion of the gbpA regulatory region |
| GBPA004 | GAGCTCTCACACTTTTTCGAGAAATTA | |
| GBPA005 | GAGCTC ACATCTATAAATAACGCTTCTAAAT | |
| GBPA006 | GAGCTCTTATGCCTGACATCACAC | |
| GBPA007 | ACTAGTCACCATTTTCCACTGCAG | |
| For primer extension analysis, EMSA, and DNase I protection assay | ||
| GBPA05-F | ATTGCCATAGCTGGTGGTTTTCA | Amplification of the gbpA upstream region |
| GBPA05-R | CCCCGCTATCTTGGGTATGGTAAAAA | Amplification of the gbpA upstream region, Extension of the gbpA transcript |
| For qRT-PCR | ||
| GBPA_qRT-F | TGAAAGCCTGGGGTGAAGCA | Quantification of the gbpA expression |
| GBPA_qRT-R | ATCGCGTAGCGTTGAGAGCG | |
a The oligonucleotides were designed using the V. vulnificus MO6-24/O genomic sequence (GenBankTM accession number CP002469 and CP002470, www.ncbi.nlm.nih.gov).
b Regions of oligonucleotides not complementary to the corresponding genes are underlined.