TABLE 1.
Bacterial and yeast strains, cloning vectors, and recombinant plasmids used in this study
| Strain or plasmid | Relevant genotype or properties | Source or reference |
|---|---|---|
| Strains | ||
| E. coli | ||
| DH5α | supE44 ΔlacU169 (φ80lacZΔM15) hsdR17 recA1 gyrA96 thi-1 relA1 | Bethesda Research Laboratories Inc., Gaithersburg, MD |
| S17-1 | Smr SprhsdR (RP4-2 kan::Tn7 tet::Mu, integrated in the chromosome) | 54 |
| BL21(DE3) | hsdS gal (λcIts857 ind1 Sam7 nin5 lacUV5-T7 gene 1) | 55 |
| B. japonicum | ||
| 110spc4 | Spr wild type | 51 |
| GEM4 | Sprfur::Ω (parental strain: USDA I110) | 24 |
| S. cerevisiae | ||
| YM4271 | MATaura3-52 his3-200 ade2-101 lys2-801 leu2-3,112 trp1-901 tyr1-501 gal4-Δ512 gal80-Δ538 ade5::hisG | 36 |
| 8730 | ICEhmu chromosomally integrated in YM4271 | This work |
| 8732 | ICEhmumut chromosomally integrated in YM4271 | This work |
| 8733 | ICE6680 chromosomally integrated in YM4271 | This work |
| 8734 | ICE7895 chromosomally integrated in YM4271 | This work |
| 8765 | Fur box chromosomally integrated in YM4271 | This work |
| Plasmids | ||
| pGEM-T Easy | Apr TA cloning vector | Promega, Madison, WI |
| pME3535XhoI | Tcr pME3535 derivative with XhoI site (Klenow fill-in of StuI site, addition of XhoI linker) | 64 |
| pASK-IBA3+ | Apr expression vector, Tet repressor, tetA-promoter/operator, C-terminal Strep-tag II | IBA, Göttingen, Germany |
| pRK290X | Tcr pRK290 derivative with XhoI site | 1 |
| pRKPolScaI | Tcr pRK290 derivative with ScaI linker | Laboratory collection |
| pNM482 | Apr pUC8 ′lacZ | 41 |
| pSUP482 | Tcr ′lacZ part from pNM482 in pSUP202pol4 | Laboratory collection |
| pUC18/19 | Apr high-copy-number cloning vectors | 45 |
| pGAD424 | Apr expressing the GAL4 activation domain (AD) | Clontech, Palo Alto, CA |
| pLacZi | Apr for integration at the URA3 locus; carries the lacZ reporter gene | Clontech, Palo Alto, CA |
| pRJ2562 | Apr (pGEM-T Easy) 1,301-bp PCR fragment comprising bll6680 and upstream region | This work |
| pRJ2563 | Apr (pGEM-T Easy) 1,306-bp PCR fragment comprising blr7895 and upstream region | This work |
| pRJ8710 | Apr (pUC18) with 1,852-bp EcoRI/BamHI fragment comprising the hmuT-hmuR intergenic region and hmuR | This work |
| pRJ8722 | Tcr (pRK290X) with 5,538-bp hmuR-lacZ fusion | This work |
| pRJ8723 | Tcr (pRK290X) with 5,521-bp hmuR-lacZ fusion | This work |
| pRJ8724 | Tcr (pRK290X) with 5,505-bp hmuR-lacZ fusion | This work |
| pRJ8730 | Apr (pLacZi) triple tandem copy of ICEhmu | This work |
| pRJ8732 | Apr (pLacZi) triple tandem copy of ICEhmumut | This work |
| pRJ8733 | Apr (pLacZi) triple tandem copy of ICE6680 | This work |
| pRJ8734 | Apr (pLacZi) triple tandem copy of ICE7895 | This work |
| pRJ8735 | Apr (pGAD424) 493-bp EcoRI fragment comprising the fur gene | This work |
| pRJ8737 | Apr (pGAD424) 504-bp SmaI fragment comprising the irr gene | This work |
| pRJ8750 | Apr (pASK-IBA3+) 491-bp Eco31I fragment comprising the irr gene | This work |
| pRJ8765 | Apr (pLacZi) triple tandem copy of Fur box | This work |
| pRJ8766 | Apr (pGEM-T Easy) 1,017-bp PCR fragment comprising bll6680 and upstream region with &12frac;-ICE6680 | This work |
| pRJ8767 | Apr (pNM482) 726-bp EcoRI/ScaI fragment of pRJ2562 comprising bll6680′-′lacZ and upstream region | This work |
| pRJ8768 | Apr (pNM482) 730-bp EcoRI/ScaI fragment of pR8766 comprising bll6680′-′lacZ and upstream region with &12frac;-ICE6680 | This work |
| pRJ8769 | Tcr (pRKPolScaI) 5,829-bp EcoRI/StuI fragment of pR8767 | This work |
| pRJ8770 | Tcr (pRKPolScaI) 5,829-bp EcoRI/StuI fragment of pR8768 | This work |