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Bacterial strains, plasmids, and oligonucleotide primers. |
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| Strain, plasmid, or primer |
Relevant characteristic(s) or sequence (5' to 3') |
Source, reference or target |
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| Strains |
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| 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 lacIqZΔM15 Tn10]) |
Stratagene1 |
| E. coli NM522 |
Cloning host (supE thi-1 Δ(lac-proAB) Δ(mcrB-hsdSM)5 (rK- mK-) [F' proAB lacIqZΔM15]) |
Stratagene1 |
| Plasmids |
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| pQE30 |
His-tag fusion protein expression vector; Cmr, Amr |
Qiagen1 |
| pGL387 |
768-pb digested PCR fragment (wzb) cloned into the SacI-KpnI site of pQE30 |
This study |
| Primers2 |
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| 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 |
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1 ATCC (American Type Culture Collection, Manassas, VA, USA); Qiagen S.A. (Courtaboeuf, France); Stratagene (LaJolla, CA, USA). 2 Restriction sites in primers are indicated in lower case letters. | ||
LaPointe et al. BMC Biochemistry 2008 9:10 doi:10.1186/1471-2091-9-10 |
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