TABLE 3.

Oligonucleotides used in PCR-mediated mutation of the vvpE promoter

Oligonucleotidea	Oligonucleotide sequence (5′–3′)b	Locationc	Use
VVPE001	GTAGGTACCACTCAAGCTGACGAACTTGATC	−335 to −314	Construction of all pHS-lux reportersd
VVPE006	GAATCTAGACCGATACAGAAGGCAGATCGGC	392 to 413
A206/197	TCTGTGGATCCTCTCTGCGTAAAAAAGA	−196 to −180	pHS206
B206/197	AGAGAGGATCCACAGATAAATGGGACAG	−207 to −223	pHS206
A176/167	AAAGTGGATCCTCTTACTAGTGTTTATC	−166 to −150	pHS176
B176/167	GTGGAGGATCCACCTTTTTCTTTTTTAC	−177 to −193	pHS176
A136/127	AATGTGGATCCTCAAACGAAATCGCAGT	−126 to −110	pHS136
B136/127	AATGAGGATCCACATAGCGATAAACACT	−137 to −153	pHS136
A106/97	ATGTGGATCCTCTATAACATTGCGTCAG	−96 to −81	pHS106
B106/97	TAGAGGATCCACATCTCACTGCGATTTC	−107 to −122	pHS106
A56/47	AACGTGGATCCTCTGAAGTTGGCTGGTG	−46 to −30	pHS56
B56/47	TCAGAGGATCCACGTTTTTACGCCCTGT	−57 to −73	pHS56
AΔ	TATAACATTGCGTCAGAGTTTCTATAC	−96 to −70	pHS221 and pHS222
BΔ−101/−97	TGACGCAATGTTATAAATCCATCTCACTGC	−82 to −116 (Δ −101 to −97)	pHS221
BΔ−106/−97	TGACGCAATGTTATAATCTCACTGCGATTTC	−82 to −127 (Δ −106 to −97)	pHS222

^a The oligonucleotides were designed using the genomic sequence of V. vulnificus CMCP6 (GenBank^TM accession numbers AE016795 and AE016796).

^b Regions of oligonucleotides not complementary to corresponding genes are underlined.

^c Shown are the oligonucleotide positions, where +1 is the transcription start site of PS.

^d Constructs are depicted in Figs. 3A and 4A.