TABLE 1.
Bacterial strains, plasmids, and primers used in this study
| Strain, plasmid, or primer | Relevant characteristica | Source or referenceb |
|---|---|---|
| Strains | ||
| A. mimigardefordensis strain DPN7T | Wild type, DTDP-degrading | 42; DSM 17166; LMG 22922 |
| Tn5::mob-induced mutants of A. mimigardefordensis (JHW51c, JHW90a, and JHW101b) | No growth with DTDP as the sole carbon source; insertion of Tn5::mob in lpdAAm | 41 |
| R. eutropha H16 | Wild type, produces polythioester | DSM 428 |
| E. coli TOP10 | F−mcrA Δ(mrr-hsdRMS-mcrBC) rpsL nupG φ80lacZΔM15 ΔlacX74 deoR recA1 araD139 Δ(ara-leu)7697 galU galK endA1 | Invitrogen, Carlsbad, CA |
| E. coli One-Shot Mach 1-T1 | lacZΔM15 hsdR lacX74 recA endA tonA | Invitrogen, Carlsbad, CA |
| E. coli (DE3)BL21/pLysS | F−ompT hsdSB (rB− mB−) gal dcm (DE3)/pLysS (Cmr) | Novagen, Madison, WI |
| Plasmids | ||
| pGEM-T Easy | Apr, lacPOZ′ | Promega, Madison, WI |
| pCR2.1TOPO | pUCori lacZ, f1 ori; Kmr Apr | Invitrogen, Carlsbad, CA |
| pET23a(+) | pBR322 ori, f1 ori His6; Apr T7lac | Novagen, Madison, WI |
| Primers | ||
| Forward-lpdANdeI | CATATGAGTCAAGTAGAAATCAAGGTG | MWG-Biotech AG |
| Reverse-lpdAXhoI | CTCGAGCTTGCGTTTGACCGGAGGC | MWG-Biotech AG |
| Forward-pdhLNdeI | AAAAACATATGAGTGTGATCGAAGTCAAGGTGCC | MWG-Biotech AG |
| Reverse-pdhLXhoI | AAAAACTCGAGGCGCTTGCGCGGCGGC | MWG-Biotech AG |
a
Cmr, chloramphenicol resistance; Apr, ampicillin resistance. For E. coli genotypes, see reference 4; LMG, Laboratorium Microbiologie Universiteit Gent.
b
DSM, Deutsche Sammlung von Mikroorganismen. Start codons are in boldface type and introduced restriction sites are underlined.