Table 1 Bacterial strains and plasmids
Strain or plasmid | Relevant characteristic(s) | Source or reference |
|---|---|---|
Strains | ||
L. rhamnosus ATCC 9595 | Low EPS-producing strain (116 mg/L) | ATCC1 |
L. rhamnosus RW-9595 M | High EPS-producing strain (1611 mg/L) | Denis Roy2 |
L. lactis subsp. cremoris MG1363 | Plasmid free (Lac-Prt-) | [17] |
E. coli NM522 | Cloning host (supE thi-1 ∆(lac-proAB) ∆(mcrB–hsdSM)5 (rK– mK–) [F´ proAB lacI q Z∆M15]) | Stratagene1 |
E. coli BL21(DE3) | Expression host (B F−, ompT, hsdS (rB −, mB −), gal, dcm) | EMD Chemicals |
E. coli C41(DE3) | Expression host derived from BL21 | [18] |
Plasmids | ||
pMG36CT | Cm, 3.7 kb, pWV01 replicon | |
pMG36EB | pMG36CT containing 1.6 kb XbaI-AatII PCR amplicon (wze and wzb) from RW-9595 M | This study |
pDWTEB | pMG36EB containing 923 bp SacI-XbaI PCR amplicon (wild type wzd) from RW-9595 M | This study |
pDY33FEB | pMG36EB containing 923 bp SacI-XbaI PCR amplicon (Y33F mutated wzd) from RW-9595 M | This study |
pDY44FEB | pMG36EB containing 923 bp SacI-XbaI PCR amplicon (Y44F mutated wzd) from RW-9595 M | This study |
pDY77FEB | pMG36EB containing 923 bp SacI-XbaI PCR amplicon (Y77F mutated wzd) from RW-9595 M | This study |
pDY110FEB | pMG36EB containing 923 bp SacI-XbaI PCR amplicon (Y110F mutated wzd) from RW-9595 M | This study |
pDY114FEB | pMG36EB containing 923 bp SacI-XbaI PCR amplicon (Y114F mutated wzd) from RW-9595 M | This study |
pDY124FEB | pMG36EB containing 923 bp SacI-XbaI PCR amplicon (Y124F mutated wzd) from RW-9595 M | This study |
pDY134FEB | pMG36EB containing 923 bp SacI-XbaI PCR amplicon (Y134F mutated wzd) from RW-9595 M | This study |
pDY141FEB | pMG36EB containing 923 bp SacI-XbaI PCR amplicon (Y141F mutated wzd) from RW-9595 M | This study |
pDY266FEB | pMG36EB containing 923 bp SacI-XbaI PCR amplicon (Y266F mutated wzd) from RW-9595 M | This study |
pQE30 | His-tag fusion protein expression vector; Cmr, Amr | Qiagen1 |
pQE31 | His-tag fusion protein expression vector; Cmr, Amr | Qiagen1 |
pGEX-6P-3 | GST fusion protein expression vector | Amersham Biosciences1 |
pGSTWze | 779 bp digested PCR fragment (wze) cloned into the BamHI-XhoI site of pGEX-6P-3 | GST-tag Wze; This study |
pQEWzd | 912 bp BamHI/XhoI digested PCR fragment (wzd) cloned into the BamHI-SalI site of pQE30 | His-tag Wzd; This study |
pQEWze | 819 bp digested PCR fragment (wze) cloned into the BamHI-HindIII site of pQE31 | native Wze; This study |
pQEWzb | 805 bp SstI-KpnI digested fragment (wzb) from pCRB cloned into the SstI-KpnI site of pQE31 | native Wzb; This study |