Table 1 Bacterial strains and plasmids used in this study
Strain or plasmid | Genotype or description | Source or references |
|---|---|---|
E. coli strains | ||
XL1-Blue | recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F’ proAB lacI qZ∆M15 Tn10 (Tetr)] | Stratagene |
BL21 (DE3) | F− ompT hsdS B (rB− mB−) gal dcm (DE3) | Novagen |
SHuffle® T7 express | fhuA2 lacZ::T7 gene1 [lon] ompT ahpC gal λatt::pNEB3-r1-cDsbC (SpecR, lacI q) ΔtrxB sulA11 R(mcr-73::miniTn10–TetS)2 [dcm] R(zgb-210::Tn10–TetS) endA1 Δgor ∆(mcrC-mrr)114::IS10 | New England BioLabs |
Plasmids | ||
pTWIN1 | pBR322 derivative (Apr) encoding two mini-inteins (Ssp DnaB and Mxe Gyr) | New England BioLabs |
pMCS69 | pBBR1MCS derivative encoding PhaA and PhaB | [15] |
pMCS69E | pBBR1MCS derivative encoding Erv1p in addition to PhaA and PhaB | [22] |
pET14b-ZZ(−)PhaC | pET14b derivative encoding ZZ-PHA synthase | [21] |
pET14b-PhaC-linker-ZZ | pET-14b derivative encoding PhaC-linker-ZZ | [16] |
pET14b-PhaC-linker-GFP | pET14b derivative encoding PHA synthase-linker-GFP | [17] |
pPOLY-C-phaC-rv1626 | pET14b derivative encoding PHA synthase-RV1626 | [20] |
pET14b-PhaC-intein-GFP | pET14b derivative encoding PHA synthase-intein-GFP | This study |
pET14b-PhaC-intein-Rv1626 | pET14b derivative encoding PHA synthase-intein-Rv1626 | This study |
pET14b-PhaC-intein-ZZ | pET14b derivative encoding PHA synthase-intein-ZZ | This study |
pET14b-PhaC-intein-TNFα | pET14b derivative encoding PHA synthase-intein-TNFα | This study |
pET14b-PhaC-intein-IFNα2b | pET14b derivative encoding PHA synthase-intein-IFNα2b | This study |
pET14b-PhaC-intein-G-CSF | pET14b derivative encoding PHA synthase-intein-GCSF | This study |