Screening of Anti-Adhesion Agents for Pathogenic Escherichia coli O157:H7 by Targeting the GrlA Activator

Table 1

Bacterial strains and plasmids used in this study.

Strain or plasmid	Relevant characteristics^a	Reference or source
Bacterial strain
E. coli
EDL933 (ATCC 43895)	Wild type, clinical isolate, virulent	Laboratory collection
EDL933 ΔgrlA	EDL933 with grlA deletion mutation	Seoul Nat’l Univ.
EDL933 grlA*	EDL933 with K111A, R114A mutations introduced into chromosomal grlA	This study
DH5α	E. coli K-12 strain for cloning experiments	Laboratory collection
SY2102	DH5α containing pSY2101 and pSY2104; Ap^r, Km^r, Cm^r	This study
SY2110	EDL933 containing pSY2104; Cm^r	This study
SY2111	EDL933 ΔgrlA containing pSY2104; Cm^r	This study
SY2201	DH5α containing pSY2201 and pSY2104; Ap^r, Km^r, Cm^r	This study
SY2204	EDL933 grlA* containing pSY2104; Cm^r	This study
Plasmids
pJK1113	pBAD24 with oriT of RP4 and nptI, P_BAD; Ap^r, Km^r	[40]
pBBR_lux	Broad host range vector with promoterless luxCDABE; Cm^r	[41]
pSY2101	pJK1113 with grlA; Ap^r, Km^r	This study
pSY2104	pBBR_lux with a fragment of LEE1/ler upstream region; Cm^r	This study
pSY2201	pJK1113 with grlA*; Ap^r, Km^r	This study
pBluescript SK II (+)	pUC ori, lacZ; Ap^r	Agilent
pCas	repA101(Ts) kan P_cas-cas9 P_araB-Red lacI^q P_trc-sgRNA-pMB1	[42]
pTargetF	pMB1 sgRNA_pMB1; Str^r	[42]
pTargetF_grlA	pMB1 sgRNA_grlA; Str^r	This study

^aAp^r, ampicillin resistant; Km^r, kanamycin resistant; Cm^r, chloramphenicol resistant; Str^r, streptomycin resistant.