title: Cloning and characterization of a cDNA encoding aspartate aminotransferase-P1 from Lupinus angustifolius root tips.
abstract: A root tip cDNA library, constructed in the lambda Zap II expression vector, was immunoscreened with a monoclonal antibody raised against aspartate aminotransferase-P1 from Lupinus angustifolius L. var Uniharvest. One 1452-base pair clone was isolated. The encoded protein sequence had high homology to both plant and animal aspartate aminotransferase sequences. The clone was converted to the phagemid form and expressed in Escherichia coli. The expressed protein was enzymically active and could be immunocomplexed with aspartate aminotransferase-P1-specific antibodies. The complete aspartate aminotransferase-P1 cDNA was cloned into the yeast expression vector pEMBL-yex4 and transformed into Saccharomyces cerevisiae strain BRSCS6, which possesses a mutated aspartate aminotransferase gene (the asp5 mutation). Complementation of the mutation was achieved using this construct.
authors.affiliation: Biochemistry Department, University of Otago, Dunedin, New Zealand.
authors.forename: C S
authors.initials: CS
authors.lastname: Winefield
authors.forename: B D
authors.initials: BD
authors.lastname: Reddington
authors.forename: W T
authors.initials: WT
authors.lastname: Jones
authors.forename: P H
authors.initials: PH
authors.lastname: Reynolds
authors.forename: K J
authors.initials: KJ
authors.lastname: Farnden
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001219:Aspartate Aminotransferases / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D009154:Mutation; D010944:Plants / Q000033:anatomy & histology / Q000201:enzymology / Q000235:genetics*; D011994:Recombinant Proteins; D012441:Saccharomyces cerevisiae; D017386:Sequence Homology, Amino Acid
chemical_list: D011994:Recombinant Proteins; D004247:DNA; D001219:Aspartate Aminotransferases