HrcU and HrpP are pathogenicity factors in the fire blight pathogen Erwinia amylovora required for the type III secretion of DspA/E

R. Ryan McNally; Quan Zeng; George W. Sundin

doi:10.1186/s12866-016-0702-y

Table 2 Bacterial strains and plasmids and their relevant characteristics

From: HrcU and HrpP are pathogenicity factors in the fire blight pathogen Erwinia amylovora required for the type III secretion of DspA/E

Strains & Plasmids	Relevant characteristics^a	Source or reference
Escherichia coli strain
DH5α	F- 80dlacZ, ΔM15, Δ(lacZYA-argF)U169, endA1, recA1, hsdR17(rK–mK+), deoR, thi-1, supE44, gyrA96, relA1 λ-	Invitrogen, CA, USA
Yeast strain
Saccharomyces cerevisiae AH109	MATa, trp1-901, leu2-3, 112, ura3-52, his3-200, gal4Δ, gal80Δ, LYS2 : : GAL1_UAS-GAL1_TATA-HIS3, GAL2_UAS-GAL2_TATA-ADE2, URA3 : : MEL1_UAS-MEL1_TATA-lacZ	[61]
Erwinia amylovora strains
Ea1189	Wild type	[62]
Ea1189∆hrcU	hrcU deletion mutant, Cm^R	This study
Ea1189∆hrpP	hrpP deletion mutant, Cm^R	This study
Plasmids
pBBR1-MCS3	Tc^R, broad host-range cloning vector	[54]
pGADT7	LEU2, Amp^R, Y2H activation vector	Clontech, CA, USA
pGBKT7	TRP1, Km^R, Y2H bait vector	Clontech, CA, USA
pLRT201	Amp^R, pMJH20 expressing DspE(1-737)-CyaA	[38]
pMJH20	Amp^R, pWSK29 containing codons 2 to 406 of CyaA	[63]
pRRM1	Tc^R, pBBR1-MCS3 containing hrcU	This study
pRRM2	Tc^R, pBBR1-MCS3 containing hrcU _N266A	This study
pRRM3	Amp^R, pGADT7 containing hrpP	This study
pRRM4	Amp^R, pGADT7 containing hrpJ	This study
pRRM5	Km^R, pGBKT7 containing hrcU	This study
pRRM6	Km^R, pGBKT7 containing hrcU _N266A	This study
pRRM7	Km^R, pGBKT7 containing hrcU _209-360	This study
pRRM8	Km^R, pGBKT7 containing hrcU _{209-360, N266A}	This study
pRRM9	Tc^R, pAlter-Ex1 containing hrcU	This study
pRRM10	Amp^R, pAlter-Ex1 containing hrcU _N266A	This study
pAlter-Ex1	Tc^R, mutagenesis vector	Promega, WI, USA
pKD3	Amp^R, Cm^R mutagenesis cassette template	[59]
pKD46	Amp^R, expresses λ red recombinase	[59]

^aCm^R, Tc^R, Amp^R, Km^R indicates resistance to chloramphenicol, oxytetracycline, ampicillin and kanamycin

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