A set of baculovirus transfer vectors for screening of affinity tags and parallel expression strategies.
We report a set of baculovirus transfer vectors for parallel expression of proteins in fusion with a panel of affinity tags including GST, protein A, thioredoxin, CBP, and FLAG. This suite includes vectors to generate recombinant baculovirus by homologous recombination in insect cells or using the Bac-to-Bac technology. An application of the vector suite approach to the vitamin D receptor (VDR), a protein mainly expressed as inclusion bodies in Escherichia coli, is presented. We found that expression in fusion with GST and protein A provided an efficient compromise of excellent purification with acceptable yields and costs.
Abdulrahman
Wassim
W
Institut de Génétique et de Biologie Moléculaire et Cellulaire, UMR 7104, Illkirch Cedex, France.
Uhring
Muriel
M
Kolb-Cheynel
Isabelle
I
Garnier
Jean-Marie
JM
Moras
Dino
D
Rochel
Natacha
N
Busso
Didier
D
Poterszman
Arnaud
A
D000345:Affinity Labels; D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D002460:Cell Line; D003001:Cloning, Molecular / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D002479:Inclusion Bodies / Q000235:genetics; D007313:Insecta; D011506:Proteins / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics; D014162:Transfection
D000345:Affinity Labels; D011506:Proteins; D011993:Recombinant Fusion Proteins