. Author manuscript; available in PMC: 2016 Jun 1.

Published in final edited form as: J Struct Funct Genomics. 2015 Apr 9;16(2):67–80. doi: 10.1007/s10969-015-9198-1

Table 2.

Expression vectors used in this study

Vector	Platform	Cloning	Antibiotic^a	Promotor/ Repressor	Encoded Fusion Protein^b	Tag Cleavage^c
pVP16	E. coli	Gateway	Amp	T5/lacI^q	His8-MBP-TEV-S-target	TEV
pVP56K	E. coli	Flexi Vector	Kan	T5/lacI	His8-MBP-TEV-AIA-target	TEV
pVP68K	E. coli	Flexi Vector	Kan	T5/lacI	His8-MBP-AIA-TEV-S-target	TEV
pVP65K	E. coli	Flexi Vector	Kan	T5/lacI	MBP-TVMV-His8-AIA-TEV-S-target	TVMV/TEV
pVP91A	E. coli	Flexi Vector	Amp	T5/lacI	His8-TEV-S-target	TEV
pEU-His	cell-free	Restriction	Amp	SP6	His6-X_2-4-target	None
pEU-His-FV	cell-free	Flexi Vector	Amp	SP6	His6-AIA-TEV-S-target	TEV

Selection marker. Amp, ampicillin; Kan: kanamycin.

His6 or His8, N-terminal histidine tag; MBP, maltose binding protein; TVMV, tobacco vein mottling virus protease recognition sequence ETVRFQ/S; TEV, tobacco etch virus protease recognition sequence ENLYFQ; AIA, protein sequence corresponding to the SgfI nucleotide sequence used for Flexi Vector cloning; S, PCR-derived substitution of the N-terminal Met with Ser to promote TEV protease action; X_2-4, 2 to 4 non-native residues whose identity varies depending on restriction enzymes used for cloning.

TEV protease is used for in vitro cleavage of fusion proteins; TVMV protease is used for in vivo cleavage of fusion proteins.