Table 1 Bacterial strains, plasmids, and oligonucleotide primers.
From: Characterization and site-directed mutagenesis of Wzb, an O-phosphatase from Lactobacillus rhamnosus
Strain, plasmid, or primer | Relevant characteristic(s) or sequence (5' to 3') | Source, reference or target |
|---|---|---|
Strains | ||
L. rhamnosus ATCC 9595 | Low EPS-producing (116 mg/L) | ATCC1 |
E. coli XL1-Blue | Cloning host (recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F' proAB lacIqZ ΔM15 Tn10]) | Stratagene1 |
E. coli NM522 | Cloning host (supE thi-1 Δ(lac-proAB) Δ(mcrB-hsdSM)5 (rK- mK-) [F' proAB lacIqZ ΔM15]) | Stratagene1 |
Plasmids | ||
pQE30 | His-tag fusion protein expression vector; Cmr, Amr | Qiagen1 |
pGL387 | 768-pb digested PCR fragment (wzb) cloned into the Sac I-Kpn I site of pQE30 | This study |
Primers 2 | ||
BF1SacI | gagctcATTGATGTGCATTGCCATATGTTACCGGGA | PCR of wzb |
BRSKpn | ggtaccTTAATACCGCGACAACAAACGCTTTTCAACC | PCR of wzb |
BD4NF | ATTAATGTGCATTGCCATATGTTACCGGGA | wzb-D3N forward |
BD4NR | TCCCGGTAACATATGGCAATGCACATTAAT | wzb-D3N reverse |
BFHI | ATTGATGTGGCTTGCGCTATGTTACCGGGA | wzb-H5A-H7A forward |
BRHI | TCCCGGTAACATAGCGCAAGCCACATCAAT | wzb-H5A-H7A reverse |
BC6SF | ATTGATGTGCATAGCCATATGTTACCGGGA | wzb-C6S forward |
BC6SR | TCCCGGTAACATATGGCTATGCACATCAAT | wzb-C6S reverse |
BH42AF | TGATGACGCCGGCCGCTATGAATGGCCG | wzb-H42-H43A forward |
BH42AR | CGGCCATTCATAGCGGCCGGCGTCATCA | wzb-H42-H43A reverse |
BD64NF | GTTTCAAAACGAGTTAGACCGCCGCAATATTCCA | wzb-D64N forward |
BD64NR | TGGAATATTGCGGCGGTCTAACTCGTTTTGAAAC | wzb-D64N reverse |
BR68AF | GTTTCAAGACGAGTTAGACGCCCGCAATATTCCA | wzb-R68A forward |
BR68AR | TGGAATATTGCGGGCGTCTAACTCGTCTTGAAAC | wzb-R68A reverse |
BR69AF | GTTTCAAGACGAGTTAGACCGCGCCAATATTCCA | wzb-R69A forward |
BR69AR | TGGAATATTGGCGCGGTCTAACTCGTCTTGAAAC | wzb-R69A reverse |
BC78SF | TTTCCCGAGTCAGGAAGTGCGGATTAATGGGC | wzb-C78S forward |
BC78SR | GCCCATTAATCCGCACTTCCTGACTCGGGAAA | wzb-C78S reverse |
BFH197A | CCTCTGATGCCGCTGCCTTACCGGGTCGCAGTTA | wzb-H197-H198A forward |
BRH197A | TAACTGCGACCCGGTAAGGCAGCGGCATCAGAGG | wzb-H197-H198A reverse |