Table 1.
Strains, plasmids, and oligonucleotides used in this study.
| Strains | Description | Reference |
|---|---|---|
| 3610 | An undomesticated strain of B. subtilis, capable of forming biofilms | 5 |
| DH5α | E. coli strain for molecular cloning | Invitrogen |
| PY79 | a competent laboratory strain of B. subtilis used for genetic manipulation | 67 |
| YC800 | ΔyvmC-cypX::tetR in 3610 | this study |
| LA11 | amyE::PyvmC-lacZ, chlR in 3610 | this study |
| LA20 | amyE::PyvmC-gfp, chlR in 3610 | this study |
| LA33 | ΔyvmC-cypX::tetR amyE::Pspank-yvmC-cypX, specR in 3610 | this study |
| YQ141 | amyE::PdhbA-lacZ, specR in 3610 | 33 |
| LA148 | ΔyvmC-cypX::tetR amyE::PdhbA-lacZ, specR in 3610 | this study |
| LA174 | amyE::PrecA-gfp, specR in 3610 | this study |
| LA224 | ΔyvmC-cypX::tetR amyE::PrecA-gfp, specR in 3610 | this study |
| LA233 | amyE::PrecA-gfp, specR lacA::PyvmC-mkate2, ermR in 3610 | this study |
| LA208 | ΔyvmA::kanR in 168 | BKK35090 |
| LA247 | ΔyvmA::kanR in 3610 | this study |
| LA262 | ΔyvmA::kanR. amyE::PdhbA-lacZ, specR in 3610 | this study |
| Plasmids | ||
| LA2 | pYC121, amyE::PyvmC-gfp, ampR in DH5α | this study |
| LA9 | pDG268, amyE::PyvmC-lacZ, ampR in DH5α | this study |
| LA22 | pDR111, amyE::Pspank-yvmC-cypX, ampR in DH5α | this study |
| LA72 | pYC121, amyE::PyvmC-mkate2, ampR in DH5α | this study |
| LA97 | pDG268, amyE::PkatE-lacZ, ampR in DH5α | this study |
| LA163 | pYC211, amyE::PrecA-gfp, ampR in DH5α | this study |
| LA186 | pDR183, lacA::PyvmC-mkate2, ampR in DH5α | this study |
| pDG268 | amyE integration vector with a promoter-less lacZ, specR, ampR | 68 |
| pDR111 | amyE integration vector with an IPTG-inducible promoter, specR, ampR | 69 |
| pDR183 | lacA integration vector, ermR, ampR | 70 |
| pYC121 | amyE integration vector with a promoter-less gfp, chlR, ampR | 71 |
| pYC211 | amyE integration vector with a promoter-less gfp, specR, ampR | this study |
| pYC251 | amyE integration vector with an IPTG-inducible promoter mkate2, chlR, ampR. | 37 |
| pDG1730 | amyE integration vector with ampr, specr, and ermr | 72 |
| Primers | ||
| PyvmC-F (for pYC121 fusion) | 5’ GTACGAATTCGCGTGTAAAAAGTGTGTGGAAAATGTG 3’ | |
| PyvmC-R (for pYC121 fusion) | 5’ GTACAAGCTTCTCATTCACCCCTAAAACTTATCCCGC 3’ | |
| PyvmC-F (for pDG268 fusion) | 5’ GTACGAATTCGCGTGTAAAAAGTGTGTGGAAAATGTG 3’ | |
| PyvmC-R (for pDG268 fusion) | 5’ GTACGGATCCCTCATTCACCCCTAAAACTTATCCCGC 3’ | |
| yvmC-F | 5’ GTACGCTAGCAAGGAGGAACTACTATGACCGGAATGGTAACGG 3’ | |
| cypX-R | 5’ GTACGCATGCTTATGCCCCGTCAAACGCAACG 3’ | |
| mkate2-F | 5’ GTACGCATGCATGGATTCAATAGAAAAGGTAAGCG 3’ | |
| mkate2-R | 5’ ACTGGTCTGATCGGATCCTTAT 3’ | |
| PrecA-F | 5’ GTACGAATTCTCGGCATTTCCGCAAATGG 3’ | |
| PrecA-R | 5’ GTACGTCGACTCTATTTTTTCCTCCTTTATG 3’ | |
| yvmC-P1 | 5’ AACAGTGTATAAGATCAATACTGCCTTTGATTTTGTATGA 3’ | |
| yvmC-P2 | 5’ CAATTCGCCCTATAGTGAGTCGTCTCATTCACCCCTAAAACTT 3’ | |
| cypX-P1 | 5’ CCAGCTTTTGTTCCCTTTAGTGAGTAGAATTCCAAAGGTCTCTCCC 3’ | |
| cypX-P2 | 5’ TTTGTGCCAAAAACTTGCGGACCGCTCCGGGGGTCGGGTC 3’ | |