Table 1.
Protein | Affinity chromatography ligand | Elution conditions | Expression system | Production yield (mg/l)a |
AVR2 | D-biotin | 0.5 M acetic acid | BEVSb | 0.8 |
AVR2-b | D-biotin | 0.5 M acetic acid | E. colic | 4.9 |
AVR2(I109K) | D-biotin | 0.5 M acetic acid | BEVS | 9.0 |
AVR4d | 2-iminobiotin | 50 mM Na-Ac + 100 mM NaCl | BEVS | 6.1 |
AVR4-bd | 2-iminobiotin | 50 mM Na-Ac + 100 mM NaCl | E. coli | 21.9 |
AVR4(K109I)d | 2-iminobiotin | 50 mM Na-Ac + 100 mM NaCl | BEVS | 16.2 |
AVR4(K109I)-bd | 2-iminobiotin | 50 mM Na-Ac + 100 mM NaCl | E. coli | 6.7 |
AVR6-be | D-biotin | 2 M acetic acid | E. coli | 8.3 |
AVD(K111I) | 2-iminobiotin | 50 mM Na-Ac + 100 mM NaCl | BEVS | 11.5 |
AVD(K111I)-b | 2-iminobiotin | 50 mM Na-Ac + 100 mM NaCl | E. coli | 0.7 |
aAfter affinity chromatography per one liter of culture medium. The yields are calculated based on only a few protein purifications.
bProduced using a baculovirus expression system in insect cells as described previously [40].
cThe bacterial signal peptide from the protein OmpA was utilised in order to produce the protein in an active form in E. coli as described in [37].
dAVR4 is identical to AVR5. The protein carries the mutation C122S, which prevents oligomerisation via intermolecular disulphide bridges [36].
eCarries mutation C58S, which prevents oligomerisation via intermolecular disulphide bridges (this study).