TABLE 1.
Bacterial strains, plasmids, and oligonucleotides used in this study
| Bacterial strain, plasmid, or primer | Genotype or sequence (5′-3′)a | Reference or source |
|---|---|---|
| Strains | ||
| E. coli DH5α | F−recA1 endA2 gyrA96 thi-1 hsdR17(rK− mK+) sup44 relA1 λ− (φ80 ΔlacZΔM15) Δ(lacZYA-argF)U169 | 19 |
| A. woodii | Wild type | DSM 1030 (DSMZ, Braunschweig, Germany) |
| Plasmids | ||
| pCR 2.1-TOPO | Invitrogen, Karlsruhe, Germany | |
| pMal-c2 | NEB, Frankfurt/Main, Germany | |
| Oligonucleotide primers | ||
| etfAforward | TGGGTNTTYGCNGARCARMGN | |
| etfAreverse | RTCNCCNACRATNCCRTARTC | |
| etfBforward | ATGGGNGCNGAYGARGCNTAY | |
| etfBreverse | NACRTANGTNACNACNGGCAT | |
| etfAforward BamHI | GTATTTGCCGGATCCCGTGAAG | |
| etfAreverse SalI | GTCGCCTACGTCGACGTAATC | |
| etfBforward EcoRI | GGAATTCATGGGGGCGGACGAAG | |
| etfBreverse PstI | AACTGCAGCGGTGGTCC | |
| iPCR_rev | GACAACCTTTTCAGCTTGCG | |
| iPCR_for | CCGGTGGACTATTGATGGG | |
| C2_for | GAATGCGAACCTTACTTAACT | |
| C2_rev | TCCCATCATAGGTCCACC |
a
In the primer sequences restriction sites introduced by PCR are underlined. N = A, C, G, or T; M = A or C; R = A or G; Y = C or T.