Strains |
|
|
B. bacilliformis
|
|
|
JB584 |
Transformation-competent strain of B. bacilliformis
|
5 |
SC1 |
JB584 with ialB interrupted by pSAC100 (Kmr, ialB mutant) |
This study |
SC2 |
SC1 complemented in trans with pSAC200 (Kmr Cmr, ialB+) |
This study |
E. coli
|
|
|
DH5α |
Host strain for cloning and plasmid propagation |
Gibco-BRL |
M15 |
Host strain for fusion protein expression |
41 |
Plasmids |
|
|
pIAL1 |
pUC19 containing ialA and ialB of B. bacilliformis
|
27 |
pIALB |
pUC19 containing ialB of B. bacilliformis
|
27 |
pQE-31 |
Expression vector |
Qiagen Inc. |
pREP4 |
Plasmid encoding lacI
|
Qiagen Inc. |
pQIALB |
pQE-31 with 574-bp PvuII-PstI fragment encoding ialB minus its secretory signal sequence plus 15 nucleotides |
This study |
pUB1 |
B. bacilliformis suicide plasmid; Kmr
|
5 |
pSAC100 |
pUB1 with an internal 430-bp PvuII-MfeI fragment of ialB; Kmr
|
This study |
pBBR1MCS |
B. bacilliformis shuttle vector; Cmr
|
18 |
pSAC200 |
Complementation plasmid; pBBR1MCS with 756-bp SwaI-BamHI fragment containing intact ialB; Cmr
|
This study |