Characterization of phosphinothricin acetyltransferase and C-terminal enzymatically active fusion proteins.
Enhanced expression of the bialaphos resistance (bar) from Streptomyces hygroscopicus, which confers resistance to the herbicides bialaphos and phosphinothricin (PPT), has been obtained in Escherichia coli using a vector system based on translational coupling. The gene product, PPT acetyltransferase, was purified to homogeneity and its enzymatic properties were analyzed. Hybrid gene constructs with gene fragments fused to the 3'-terminus of bar yield fusion proteins having acetyltransferase activity, with a Michaelis constant for the PPT substrate comparable to the unmodified enzyme. The bar gene represents a selectable and assayable reporter gene especially suitable for 3'-terminal gene fusions.
Botterman
J
J
Plant Genetic Systems N.V. Gent, Belgium.
Gosselé
V
V
Thoen
C
C
Lauwereys
M
M
D000123:Acetyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000613:Aminobutyrates / Q000378:metabolism* / Q000494:pharmacology; D015153:Blotting, Western; D004926:Escherichia coli / Q000378:metabolism; D015870:Gene Expression; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009943:Organophosphorus Compounds / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry; D013302:Streptomyces / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D013696:Temperature
D000613:Aminobutyrates; D009943:Organophosphorus Compounds; D011993:Recombinant Fusion Proteins; C003122:bialaphos; C003121:phosphinothricin; D000123:Acetyltransferases; C056243:phosphinothricin N-acetyltransferase