TABLE 2.

Oligonucleotide primers

Primera	sequence (5′ - 3′)b	Usec
vca1008F1	CAGGACGCAATGGAGTAGTC	SOE PCR
vca1008F2	GGTAGAAAAATGTAATCACACTGCCTTAAAC	SOE PCR
vca1008R1	AGTGTGATTACATTTTTCTACCCTATTAG	SOE PCR
vca1008R2	TGGCGTGATCAAACGGCTGGAC	SOE PCR
ompUF1	CAGCATGGTATTCCGCATTC	SOE PCR
ompUF2	ATGGACAATACTTCAGGTCACACGCCAAAC	SOE PCR
ompUR1	GACCTGAAGTATTGTCCATAAATTTG	SOE PCR
ompUR2	GATCAGGTTGTCGGACTCTTG	SOE PCR
ompTF1	TGATCACTGATCCTGCGA	SOE PCR
ompTR1	GATCTTACAACTCTTTGTTTGGTCACCAC	SOE PCR
ompTF2	CAAAGAGTTGTAAGATCTCGAACACGTTTA	SOE PCR
ompTR2	CATCCCTCTTGCCAAGCCAG	SOE PCR
vc0972F1	GGTAGACTCTGCTCTAGC	SOE PCR
vc0972R1	CGTCTGATTACATGGATAACTCCTAAAAATG	SOE PCR
vc0972F2	GTTATCCATGTAATCAGACGAACCCTGC	SOE PCR
vc0972R2	AAGAGTGAGCATGGCTTTAG	SOE PCR
vca1008F0	CAAGCAATTAAATTGCACAC	Deletion confirmation
ompUF0	TAGCTTGTATTCGATATCAC	Deletion confirmation
ompUR0	AGTAAAAACGGTGTCCCAAGG	Deletion confirmation
ompTF0	ACGTTCGCTACAACAATAAC	Deletion confirmation
vc0972F0	ACCGTAACCAACAAGTGATC	Deletion confirmation
GOA3	TCTAATGAATTCTGCGTGGCAACATTGATGTG	vca1008 cloning
GOA4	TCTAATTCTAGAGGTGATGTTTAATGCTCATG	vca1008 cloning
GOA5	TCTAATTCTAGATGCGTGGCAACATTGATGTG	vca1008 cloning
GOA6	TCTAATGAATTCGGTGATGTTTAATGCTCATG	vca1008 cloning

^aThe beginning of each primer name corresponds to the V. cholerae N16961 gene being deleted, and the F and R indicate forward and reverse primers, respectively.

^bUnderlined bases indicate a noncomplementary 5′ tail and restriction site.

^cSOE PCR refers to splicing-by-overlap-extension PCR used to construct gene deletions.