E. faecalis strain(s) |
|
|
OG1RF |
Used for lsa insertional mutagenesis and to amplify lsa for complementation experiments; Rifr, Fusr
|
14 |
V583 |
TIGR-sequenced strain; used to amplify lsa for complementation experiments |
17, 22
|
TX4107-TX4110 |
Q-D- and CLI-susceptible isolates from animal feed |
This study |
TX0263, TX0271 |
Q-D- and CLI-susceptible clinical isolates |
This study |
TX5076 |
Antigen gene disruption mutant generated with pTEX4577 containing aph(3′)-llla; Kanr
|
30 |
TX5243 |
sprE gene disruption mutant [OG1RF sprE::pTEX4577 containing aph(3′)-llla]; Kanr
|
18 |
TX5248 |
orf1 gene disruption mutant [OG1RF orf1::pTEX4577 containing aph(3′)-llla]; Kanr
|
18 |
TX5332 |
lsa gene disruption mutant [OG1RF lsa::pTEX4577 containing aph(3′)-llla]; Kanr
|
25 |
|
TX5332(pTEX5333.04) (contains lsaV); Kanr, Chlr
|
25 |
|
TX5332(pTEX5333.03) (contains lsaOG); Kanr, Chlr
|
This study |
|
TX5332(pTEX5333.08) (contains lsaOG5′V3′); Kanr, Chlr
|
This study |
|
TX5332(pTEX5333.09) (contains lsaV5′OG3′); Kanr, Chlr
|
This study |
|
TX5332(pTEX5333.12) (contains a mutagenized gene, lsaV-M); Kanr, Chlr
|
This study |
|
TX5332(pTEX5333.13) (contains a mutagenized gene, lsaOG-M); Kanr, Chlr
|
This study |
L. lactis strains |
|
|
LM2301 |
Used as a gram-positive host for E. faecalis lsa
|
27 |
|
LM2301(pTEX5333.04) (contains lsaV5); Chlr
|
This study |
|
LM2301(pTEX5333.03) (contains lsaOG); Chlr
|
This study |
|
LM2301(pTEX5333.08) (contains lsaOG5′V3′); Chlr
|
This study |
|
LM2301(pTEX5333.09) (contains lsaV5′OG3′); Chlr
|
This study |
|
LM2301(pTEX5333.12) (contains lsaV-M); Chlr
|
This study |
|
LM2301(pTEX5333.13) (contains lsaOG-M); Chlr
|
This study |
TX5392 |
LM2301(pWM401); Chlr
|
This study |
Plasmids |
|
|
pTEX4577 |
pBluescript SK(−) with aph(3′)-IIIa inserted into the ScaI site; Kanr, used for insertional mutagenesis |
24 |
pWM401 |
Shuttle vector; Chlr, Tetr
|
29 |
pCR2.1 vector |
PCR product cloning vector |
Invitrogen |
pTEX5333.03 |
pWM401::lsaOG, contains a ca. 2-kb lsa fragment from OG1RF, used for complementation of TX5332 and transformation of L. lactis LM2301; Chlr
|
This study |
pTEX5333.04 |
pWM401::lsav, contains a ca. 2-kb lsa fragment from V583, used for complementation of TX5332 and transformation of L. lactis LM2301; Chlr
|
25 |
pTEX5333.08 |
pWM401::lsaOG5′V3′, contains a hybrid gene with the 5′ half of lsaOG and 3′ half of lsav, used for complementation of TX5332 and transformation of L. lactis LM2301; Chlr
|
This study |
pTEX5333.09 |
pWM401::lsaV5′OG3′, contains a hybrid gene with the 5′ half of lsaV and 3′ half of lsaOG, used for complementation of TX5332 and transformation of L. lactis LM2301; Chlr
|
This study |
pTEX5333.12 |
pWM401::lsaV-M, contains ca. 2-kb lsaV fragment mutagenized in its upstream region at positions −131 and −133 with A's replaced with a G and a T, used for complementation of TX5332 and transformation of L. lactis LM2301; Chlr
|
This study |
pTEX5333.13 |
pWM401::lsaOG-M, contains ca. 2-kb lsaOG fragment mutagenized in its upstream region at positions −131 and −133 with G and T replaced by A's, four additional nucleotide changes were found at −109, +1248 (silent), +1572 (downstream), and +1517 (downstream), used for complementation of TX5332 and transformation of L. lactis LM2301; Chlr
|
This study |