Lack of a type-2 glycosyltransferase in the fish pathogen Flavobacterium psychrophilum determines pleiotropic changes and loss of virulence

Table 1 Bacterial strains, plasmids and primers used in this study

Plasmid, bacterial strain or primer	Description or sequence	Source or reference
Plasmids^a
pEP4351	Ori R6K dependent protein pir,; RP4 oriT; Cm^r Tc^r (Em^r); Tn4351 vector transfer.	[39]
pCP23	ColE1 ori, (pCP1 ori), Ap^r (Tc^r), E. coli–F. psychrophilum shuttle plasmid	[40]
pCP23-fpgA	FP1638 gene, derived from pCP23.	This study
pCP23-Gfpp2	pCP23-G carrying fpp2-fpp1 promoter	[41]
Bacterial strains
F. psychrophilum
THC02/90	Wild-type.	[42]
fpgA⁻	FP1638 mutant created by Tn4351 transposition.	This study
fpgA+	Mutant ΔFP1638 carrying pCP23-FP1638 plasmid, complemented strain.	This study
THC02/90-G	THC02/90 strain carrying pCP23-G plasmid	[41]
THC02/90 -fpp2	THC02/90 strain carrying pCP23-Gfpp2 plasmid	[41]
fpgA⁻ -fpp2	ΔFP1638 strain carrying pCP23-Gfpp2 plasmid	[41]
E. coli
S17-1 λpir	λpir hsdR pro thi; RP4-2 Tc::Mu Km::Tn7	[34]
BW19851	RP4-2tet::Mu-1kan::Tn7 integrant; _uidA::pir_ recA1 hsdR17 creB510 endA1 zbf-5 thi	[35]
Primers^b
26-F	5′ACTGGGATCCAGTTTTAAGCCCGCAAA 3′	This study
26-R	5′ ACTGCTGCAGCAATGAACTTCGTCTTG 3′	This study
TN-1	5′ GGACCTACCTCATAGACAA 3′	[19]
IS4351-F	5′ TCAGAGTGAGAGAAAGGG 3′	[19]
promfpp2-F	5′ ATCAGGATCCGAGCACTACACTTTCTAGA 3′	[41]
promfpp2-R	5′ GATTGGATCCTGTTCGGTAGTGTAGCA 3′	[41]

^aAntibiotic resistance phenotypes: ampicillin, Ap^r; tetracycline, Tc^r; erythromycin, Emr^b. Antibiotic resistance phenotypes and other features listed in parentheses are those expressed by F. psychrophilum but not by E. coli.
^bRestriction sites for cloning are in bold.

ISSN: 1297-9716