Table 1 Strains, plasmids, and primers used in this study
Strain, plasmid or primer | Characteristics, or sequence | Source or reference |
|---|---|---|
Strains | ||
E. coli JM109 | recA1, endA1, gyrA96, thi-1, hsdR17 (rk−mk+), e14−(mcrA−) supE44, relA1, Δ (lac-proAB)/F′[traD36, proAB+, lacIq, lacZΔM15] | Takara |
B. coagulans 2–6 | Wild strain isolated from soil, DSM21869 | 5 |
B. coagulans Na-2 | Mutant strain of B. coagulans 2–6 with high sodium lactate tolerance ability | 14 |
B. coagulans DSM1 | Type strain, transformation host | DSMZ |
B. coagulans DSM1/pNW33n | Control strain, B. coagulans DSM1 harboring pNW33n | This study |
B. coagulans DSM1/pNW33n-mfs-2-6 | Heterogeneous expression strain, B. coagulans DSM1 harboring pNW33n-mfs-2-6 | This study |
B. coagulans DSM1/pNW33n-mfs-Na-2 | Heterogeneous expression strain, B. coagulans DSM1 harboring pNW33n-mfs-Na-2 | This study |
Plasmids | ||
pNW33n | 4.2 kb, Cmr, Geobacillus-E. coli shuttle vector | 29 |
pNW33n-mfs-2-6 | Cmr, pNW33n carrying native mfs gene from B. coagulans 2–6 and its promotor | This study |
pNW33n-mfs-Na-2 | Cmr, pNW33n carrying mutated mfs gene from B. coagulans Na-2 and its promotor | This study |
Primersa | ||
MFS-F | 5′-TCTGATTGTGAAATTGAATTCAGCAAGCCCTAAGTATGTAAAATAC-3’ | This study |
MFS-R | 5′-CTGCGGTGCGGCCATGGATCCTAACGCACCTTTGTTGTCTGCT-3’ | This study |
RT-mfs-F | 5′-ACGTTGCCGAAGGTTTGACT-3’ | This study |
RT-mfs-R | 5′-AACACAGCCCAGCTTACTCC-3’ | This study |
RT-16S-F | 5′-GGGAAACCGGGGCTAATACC-3’ | This study |
RT-16S-R | 5′-TCTGTAAGTGGCAGCCGAAG-3’ | This study |