Table 1
Constructs tested for expression of different USP18 fusion proteins
Construct (promoter, antibiotic resistance) pGEMEX-His-USP18 (T7, ampicillin) pET15b-His-USP18 (T7, ampicillin) pACE-His6-SUMO-USP18 (T7, ampicillin) pACE-His6-SUMO-TFAAA-USP18 (T7, ampicillin)
strain Rosetta(DE3) Rosetta(DE3) BL21(DE3) BL21(DE3)pLysS BL21(DE3) BL21(DE3)pLysS
temperature 25°C 25°C 37°C 37°C 25°C 37°C 37°C
expression - - - + + ++ ++
soluble n.d. n.d. n.d. n.d. n.d. n.d. n.d.
Construct (promoter, antibiotic resistance) pSUMO-His-SUMO-TFAAA-USP18 (T7, kanamycin)
strain BL21(DE3) BL21(DE3)pLysS Tuner(DE3)pLysS Tuner(DE3)
temperature 25°C 37°C 15°C 25°C 37°C 15°C 15°C
expression +++ +++ + +++ +++ ++ +++
soluble n.d. n.d. + - - ++ +

n.d.: not determined, -: no expression observed, +: detectable on Western Blot, but not on Coomassie-stained SDS PAGE gel, ++: detectable on Coomassie-stained gel, but not the dominant band, +++: dominant band on Coomassie-stained gel.

Basters et al. BMC Biotechnology 2012 12:56   doi:10.1186/1472-6750-12-56

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