High yield expression of catalytically active USP18 (UBP43) using a Trigger Factor fusion system

BMC Biotechnology

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-TF_AAA-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-TF _AAA -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.

ISSN: 1472-6750