TABLE 1.

Plasmids and strains used in this study

Strain or plasmid	Description	Reference or Source
Strains
M. voltae
PS	Wild-type strain
P-2	Flagellum-deficient mutant of M. voltae PS	17
E. coli
BL21(DE3)/pLysS	Host for T7-based expression vectors	45
DH5α
P. aeruginosa
PAK	Wild-type host
PAK-DΩ	Ω insertion mutation into pilD (prepilin peptidase gene)	28
PAO1 PilA−	Mutation in major pilin (PilA)	J. S. Mattick
Plasmids
pKJ41	Source of M. voltae flaB2 gene; missing 5′ 15 nucleotides	21
pAW102	contains 1.24-kb HindIII fragment which encodes P. aeruginosa pilA gene and promoter in pUCP vector	J. S. Mattick
pT7-7	T7 promoter-based expression vector	46
pKJ91	pT7-7 with flaB2 gene in NdeI site; same orientation as T7 promoter	This study
pKJ139	As above with flaB2 excised by XbaI and religated in opposite orientation	This study
pUCP18/19	E. coli-P. aeruginosa shuttle vector based on pUC18/19	37
pKJ118	pUCP18 with flaB2 gene excised from pKJ91 in XbaI site; fragment in same orientation as lac promoter	This study
pKJ119	As above with flaB2 in opposite orientation	This study
pKJ156	pUCP18 with fusion of pilA promoter, ribosome binding domain, leader peptide, and +1 Phe to flaB2 mature flagellin coding region	This study
pKJ248	pUCP19 with fusion of pilA promoter, ribosome binding domain, leader peptide, +1 Phe, and +5 Glu to flaB2 mature flagellin coding region	This study