Table 3 E. coli strains, plasmids, and primers used in this study
Strains, plasmids, or primers | Genotypes, relevant characteristics, or sequences | Source or references |
|---|---|---|
Strains | ||
 TOP10 | F− mcrA Δ(mrr-hsdRMS-mcrBC) Ф80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara-leu) 7697 galU galK rpsL (StrR) endA1 nupG, streptomycin-resistant | Invitrogen |
 BL21(DE3) | F- ompT hsdS B(r −B m −B ) gal dcm λ(DE3), carrying the T7 RNA polymerase gene | Novagen |
 JH0 | BL21(DE3) ΔfdhF::FRT-kan-FRT | This study |
 JH1 | BL21(DE3) ΔhycE::FRT-kan-FRT | This study |
 MW1001 | lacI q rrnB T14 ΔlacZ WJ16 hsdR514 ΔaraBAD AH33 ΔrhaBAD LD78 ΔhyaB ΔhybC ΔhycE | [33] |
Plasmids | ||
 pGEM-T Easy | bla lacZ, TA cloning vector | Promega |
 pEMBTL-HJ2 | Expression vector with T7 promoter carrying R. sphaeroides hupS and hupL | [18] |
 pTrc-EcH1ABHis | Expression vector with trc promoter carrying E. coli hyaB and hyaA | [19] |
 pTrcHis C | pBR322 ori bla lacI q, a parental expression vector with trc promoter | Invitrogen |
 pTrcHoxGK | pTrcHis C carrying H. marinus hoxG and hoxK | This study |
 pTrcHoxG | pTrcHis C carrying H. marinus hoxG | This study |
 pTrcHoxK | pTrcHis C carrying H. marinus hoxK | This study |
 pTrcHoxGK* | pTrcHis C carrying H. marinus hoxG and hoxK without signal sequence | This study |
 pTrcHoxK* | pTrcHis C carrying H. marinus hoxK without signal sequence | This study |
 pKD46 | bla γ β exo araC, Red recombinase vector containing temperature-sensitive replicon | CGSC |
 pKD13 | bla FRT-kan-FRT, template plasmid used for Red recombination | CGSC |
Primersa | ||
 hoxG | Forward: GCTAGC ATGAGCGTATTAAACACACC (NheI) Reverse: CTCGAG TTATCGAACCTTGACGGT (XhoI) | This study |
 hoxK_poly | Forward: CTCGAGTCTGCCCGTATTGCGCGTAAGGAAATCCATTATGTCAT CTCAAGTTGAAAC (XhoI) Reverse: CTGCAGTCAATGGTGATGGTGATGATGACCGCCTTTATCTCCTT TCTTTTGAGCC (PstI) | This study |
 hoxK*_poly | Forward: CTCGAGTCTGCCCGTATTGCGCGTAAGGAAATCCATTATGGCG AACAAAATTGCTCATGCGAT (XhoI) Reverse: ditto | This study |
 hoxK | Forward: CCATGGGCTCATCTCAAGTTGAAACGTT (NcoI) Reverse: ditto | This study |
 hoxK* | Forward: CCATGGGCAACAAAATTGCTCATGCGAT (NcoI) Reverse: ditto | This study |
 fdhF13 | Forward: CAATCACGTACTGCTCGGCGGCGCGCTGATCGGCGATCGGCTCG ACGCGCATTCCGGGGATCCGTCGACC Reverse: TCCTGACCCCGCGCCTGAAAACCCCCATGATCCGTCGCCAGCGT GGCGGCTGTAGGCTGGAGCTGCTTCG | This study |
 hycE13 | Forward: TTTTTGATAAAGGTAAACATGGCGATTCCTTATTTCAGCGGCGA GTTTTTATTCCGGGGATCCGTCGACC Reverse: TTAGCGTTCGTCTCCTTGCTGGCGGCGTGATTAAAGAGAGTTTG AGCATGTGTAGGCTGGAGCTGCTTCG | This study |
 fdhFchk | Forward: GTAGGGAGTAACCAGTATAA Reverse: AATGACCCCACATAAAATGT | This study |
 hycEchk | Forward: CCAGCGGATAAGACGAGGT Reverse: CGTCTTGATATTACTCCGCG | This study |