title: Expression of poplar phenylalanine ammonia-lyase in insect cell cultures.
abstract: A cDNA encoding one of the phenylalanine ammonia-lyase genes from Populus trichocarpa x deltoides was inserted into a baculovirus expression vector and the PAL protein was successfully expressed in insect cell cultures. High levels of active holoenzyme were obtained that could be purified in a single chromatographic step. Site-directed mutagenesis and expression of the mutant enzyme confirmed that conversion of the putative active site serine202 residue to alanine is sufficient to destroy the catalytic activity of PAL.
authors.affiliation: Department of Plant Science, University of British Columbia, Vancouver, Canada.
authors.forename: G R
authors.initials: GR
authors.lastname: McKegney
authors.forename: S L
authors.initials: SL
authors.lastname: Butland
authors.forename: D
authors.initials: D
authors.lastname: Theilmann
authors.forename: B E
authors.initials: BE
authors.lastname: Ellis
mesh_terms: D000818:Animals; D016367:Baculoviridae; D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D018076:DNA, Complementary; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009838:Oligodeoxyribonucleotides; D010650:Phenylalanine Ammonia-Lyase / Q000096:biosynthesis* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D017384:Sequence Deletion; D012694:Serine; D018411:Spodoptera; D014162:Transfection; D014197:Trees / Q000201:enzymology*
chemical_list: D018076:DNA, Complementary; D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins; D012694:Serine; D010650:Phenylalanine Ammonia-Lyase