Use of translational fusions to the maltose-binding protein to produce and purify proteins in Pseudomonas syringae and assess their activity in vivo.
A simple approach is described for the production and purification of proteins in Pseudomonas syringae. The strategy involves the use of the tac promoter, the maltose-binding protein, and the broad-host-range vector, pRK415. This approach was used to partially purify two proteins involved in coronatine biosynthesis from P. syringae. The activity of the fusions was demonstrated in vivo in complementation experiments using the appropriate mutants.
Peñaloza-Vázquez
A
A
Department of Plant Pathology, Oklahoma State University, Stillwater 74078-3032, USA.
Rangaswamy
V
V
Ullrich
M
M
Bailey
A M
AM
Bender
C L
CL
D000596:Amino Acids / Q000378:metabolism; D001427:Bacterial Toxins / Q000096:biosynthesis; D002352:Carrier Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D003001:Cloning, Molecular / Q000379:methods; D007192:Indenes / Q000378:metabolism; D008320:Maltose; D058619:Maltose-Binding Proteins; D010957:Plasmids; D014176:Protein Biosynthesis; D011549:Pseudomonas; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D015183:Restriction Mapping
D000596:Amino Acids; D001427:Bacterial Toxins; D002352:Carrier Proteins; D007192:Indenes; D058619:Maltose-Binding Proteins; D011993:Recombinant Fusion Proteins; C058487:coronatine; D008320:Maltose