1. Vectors Available to Arabidopsis Community [Link]
2. IUE Virtual Laboratory Course [Link]
3. Evaluation of the affinity tag
(Evaluation will involve determining the yield and fold purification in a side by side comparison with the Arabidopsis exosome complex purification as described in Chekanova et al, Cell 131:1340-1353, 2007)
SUGGEST YOUR OWN TAG FOR TESTING
FLAG (DYKDDDDK)
Hopp et al (1988) Bio/Technology 6:1204
His (HHHHHH or HHHHHHHH or RSGHHHHHH)
Crowe et al (1994) Methods in Molecular Biology 31:371
Myc (EQKLISEEDL)
Hilpert et al (2001) Protein Engineering 14:803
HA (YPYDVPDYA)
Field et al (1988) Mol Cell Biol 8:2159
T7 (MASMTGG QQMG)
Makrides (1996) Microbiol Rev 60:512
AcV5 (SWKDASGWS)
Hohmann et al (1983) Virology 125:432, Lawrence et al (2003) Biotechniques 35:488
V5 (GKPIPNPLLGLDS T)
Southern et al (1991), J Gen Viol 72:1551
SBP (MDEKTTGWRGGHVVEGLAGELEQLR ARLEHHPQGQREP)
Keefe et al (2001) Protein Expression and Purification 23:440
S-tag (KETAAAKFERQHMDS)
Raines et al (2000) Meth Enzymol 326:362
VSV-G tag (YTDIEMNRLGK)
Le Maout et al (1997) PNAS 94:13329
BAD (MAGLNDIFEAQKIEWHE)
De Boer et al (2003) PNAS 100:7480
Protein A (32 kDa; ZZ portion is16 Da )
Uhlen et al (1983) Gene 23:369
Protein C (EDQVDPRLIDGK)
Stearns et al (1988) J Biol Chem 263:826
CBP
Stearns et al (1988) J Biol Chem 263:826
GST (26 kDa; EC 2.5.1.18)
Smith & Johnson (1988) Gene 67:31
Strep II (WSHPQFEK)
Voss & Skerra (1997) Protein Eng 10:975