Table 1 Strains, plasmids, and primers used in this study
Strains and plasmids | Relevant genotypes and descriptions | Source |
|---|---|---|
Strains | ||
E. coli DH5α | F− endA1 glnV44 thi-1 recA1 relA1 gyrA96 deoR nupG Φ80dlacZΔM15 Δ(lacZYA-argF)U169, hsdR17(r − K m + K ), λ– | Lab stock |
E. coli BL21 (DE3) | F– ompT gal dcm lon hsdSB(r − B m − B ) λ(DE3 [lacI lacUV5-T7 gene 1 ind1 sam7 nin5]) | Lab stock |
E. coli MG1655 | Genomic DNA was prepared for sequence encoding Ag43 PCR-amplification. | Lab stock |
Plasmids | ||
pET28a | KanR, f1, pBR322, PT7 | Lab stock |
pSB1C3-RFP | CmR, pMB1, PlacI, RFP | Lab stock |
pET28a-BsCel5 | KanR, f1, pBR322, PT7, BsCel5 | Lab stock |
pET28a-Ag43 | KanR, f1, pBR322, PT7, Ag43 | This study |
pET28a-Ag43-138-RFP | KanR, f1, pBR322, PT7, Ag43-138-RFP | This study |
pET28a-Ag43-700-RFP | KanR, f1, pBR322, PT7, Ag43-700-RFP | This study |
pET28a-RFP | KanR, f1, pBR322, PT7, RFP | This study |
pET28a-Ag43-138-BsCel5 | KanR, f1, pBR322, PT7, Ag43-138- BsCel5 | This study |
pET28a-Ag43-700-BsCel5 | KanR, f1, pBR322, PT7, Ag43-700- BsCel5 | This study |
Primers | Sequence (5′–3′)a |
|---|---|
Ag43-MG1655-EcoRI-F | TGGAATTCATGAAACGACATCTGAATACCTGCT |
Ag43-MG1655-HindIII-R | AGAAGCTTTCAGAAGGTCACATTCAGTGTGGCCT |
Ag43-RFP-IF | CACGTCACTCCCGGTGCTGGCTGCTATGGCTTCCTCCGAAGACGTTATC |
Ag43-RFP-138-IR | CAGCCCGTCCCTGAAGGCTCTGTCCAGCACCGGTGGAGTGACGACCTTC |
Ag43-RFP-138-VF | GAAGGTCGTCACTCCACCGGTGCTGGACAGAGCCTTCAGGGACGGGCTG |
Ag43-RFP-VR | GATAACGTCTTCGGAGGAAGCCATAGCAGCCAGCACCGGGAGTGACGTG |
Ag43-RFP-700-IR | CTGCACGATAAGCATTTTCACTGCGAGCACCGGTGGAGTGACGACCTTC |
Ag43-RFP-700-VF | GAAGGTCGTCACTCCACCGGTGCTCGCAGTGAAAATGCTTATCGTGCAG |
RFP-XhoI-F | GACTCGAGAGCACCGGTGGAGTGACGACCTT |
RFP-BamHI-R | CAGGATCCATGGCTTCCTCCGAAGACGTTATC |
Ag43-Cel5-IF | CACGTCACTCCCGGTGCTGGCTGCTATGGCAGGGACAAAAACGCCAGTAG |
Ag43-Cel5-138-IR | CAGCCCGTCCCTGAAGGCTCTGTCCATTTGGTTCTGTTCCCCAAATCAGT |
Ag43-Cel5-138-VF | ACTGATTTGGGGAACAGAACCAAATGGACAGAGCCTTCAGGGACGGGCTG |
Ag43-Cel5-VR | CTACTGGCGTTTTTGTCCCTGCCATAGCAGCCAGCACCGGGAGTGACGTG |
Ag43-Cel5-700-IR | CTGCACGATAAGCATTTTCACTGCGATTTGGTTCTGTTCCCCAAATCAGT |
Ag43-Cel5-700-VF | ACTGATTTGGGGAACAGAACCAAATCGCAGTGAAAATGCTTATCGTGCAG |