Table 2.

Sequences of the primers used for the construction of mutants and expression vectors.

Primer	Sequence (5′ to 3′)
Used for gene deletion
vscF-1	CAGGTCGACTCTAGAGGATCCGGGTTATCGCATATCCAG
vscF-2	CGTTCCATGCACTGTTGGTCGCATCGTA
vscF-3	GACCAACAGTGCATGGAACGAGAATTAC
vscF-4	CGAGCTCGGTACCCGGGGATCCTTCCTAATGTCTTGGGTG
vpa0226-1	CAGGTCGACTCTAGAGGATCCTGACCGTGATGCCAAAAT
vpa0226-2	AGCCGTGTCTGCGATACCAACAGCGAAC
vpa0226-3	TTGGTATCGCAGACACGGCTTCTGAGTT
vpa0226-4	CGAGCTCGGTACCCGGGGATCCAATGCCTTGTCGGCACGA
Used for the construction of complementation strains
C-vscF-1	CAGGTCGACTCTAGAGGATCCGGGTTATCGCATATCCAG
C-vscF-2	CGAGGTTTACACTGTTGGTCGCATCATA
C-vscF-3	GACCAACAGTGTAAACCTCGATGACGTTAA
C-vscF-4	CGAGCTCGGTACCCGGGGATCCTTCCTAATGTCTTGGGTG
S151A-1	CAGGTCGACTCTAGAGGATCCGGGATGATGAAAAAAACAATCACAC
S151A-2	GTATCAGACAATGCGTCACCGAGTGCAACC
S151A-3	GGTGACGCATTGTCTGATACAGGCAACATC
S151A-4	CGAGCTCGGTACCCGGGGATCCAATGCCTTGTCGGCACGAGA
Used for the construction of expression vectors
pM207-vp1683CyaA1	GAGCTCGGTACCCGGGGATCCCATGGTTAATATCAATACGTC
pM207-vp1683CyaA2	GCGATTGCTGACCAAGTTTGTGGCTAT
pM207-vp1683CyaA3	CAAACTTGGTCAGCAATCGCATCAGGCTGG
pM207-vp1683CyaA4	CAGGTCGACTCTAGAGGATCCTTATGTCATAGCCGGAATCC
pM207-vp1686CyaA1	GAGCTCGGTACCCGGGGATCCCATGATCAGTTTTGGAAATGT
pM207-vp1686CyaA2	GCGATTGCTGTTTGATACCGTGAAGGCTAT
pM207-vp1686CyaA3	CGGTATCAAACAGCAATCGCATCAGGCTGG
pM207-vp1686CyaA4	CAGGTCGACTCTAGAGGATCCTTATGTCATA GCCGGAATCC
pM207-vpa0226CyaA1	GAGCTCGGTACCCGGGGATCCCATGATGAAAAAAACAATCAC
pM207-vpa0226CyaA2	GCGATTGCTGGAAACGGTACTCGGCTAAGT
pM207-vpa0226CyaA3	GTACCGTTTCCAGCAATCGCATCAGGCTGG
pM207-vpa0226CyaA4	CAGGTCGACTCTAGAGGATCCTTATGTCATA GCCGGAATCC
Used for qRT-PCR
16sRNA-F	GACACGGTCCAGACTCCTAC
16sRNA-R	GGTGCTTCTTCTGTCGCTAAC
vpa0226RT-F	CAAACCAGCAAACACCTT
vpa0226RT-R	GTCCGTCAAACGAATCAG

Underscore served as indicate vector upstream or downstream sequence.