Serratia marcescens internalization and replication in human bladder epithelial cells

BMC Infectious Diseases

Table 1 Bacteria, plasmids, and epithelial cells used in this study

Strains/plasmids/cell lines	relevant genotype	source or reference
E. coli
BL21 [DE3]	F^-, hsdS gal, lysogenic for phage λ carrying the phage T7 RNA polymerase gene under lacUV5 control	[36]
SM10[λpir]	thi-1 thr leu tonA lacY supE recA::RP4-2-Tc::mu, Km^r, λpir	[28]
HB101	ara leu pro lac gal rpsL supE44 xyl mtl recA hsd9 (hsdR hsdM)	Stratagene
S. marcescens
W225	human isolate	[31]
SM001	W225 shlB/shlA::kan, nonhemolytic	this study
SM011	W225 shlB/shlA::kan, nonhemolytic	this study
VA 15854	human isolate	[31]
W1128	human isolate	[30]
ATCC25419	human isolate	[31]
CDC 04:H4	human isolate	[31]
Other strains
Proteus mirabilis VI	human isolate	[34]
Pseudomonas aeruginosa	PAO1	S. Lory
Plasmids
pGP704	Ap^r; ori R6 K, mob RP4, MCS of M13tg131	[13]
pRK2013	conjugative IncP helper plasmid	ATCC 37159
pMMB67EH	multi-host cloning and expression vector	ATCC 37622
pRSC01	pMMB67EH shlB shlA	this work
pUC4-K	Ap^r, Km^r; MCS	[37]
pRSM01	Ap^r, Km^r; ori R6 K, mob RP4, MCS of M13tg131	this work
pRSM02	pRSM01, 1.8-kb SphI fragment of pRO2 [shlB'/shlA']	this work
pRSM03	pRSM02, SgrAI deletion [shlB/shlA, Δ bp 1544–2285]	this work
pRO3	pT7-5 shlB shlA	[28]
pRO2	pT7-5 shlB shlA, Δ aa 256–1578	[28]
Epithelial cells
RT112	human bladder epithelial carcinoma cell line	MPI Tübingen
HEp-2	human larynx epidermoid carcinoma cell line	ATCC CCL23
HeLa	human cervix epitheloid carcinoma cell line	ATCC CCL 2
Hec1-B	human endometrial adenocarcinoma cell line	ATCC HTB 113

ISSN: 1471-2334