title: Cloning and expression of a functional rat liver D-beta-hydroxybutyrate dehydrogenase-beta-galactosidase fusion protein in Escherichia coli.
abstract: A rat liver bacteriophage lambda expression library was probed using polyclonal antibodies raised to purified rat liver D-beta-hydroxybutyrate dehydrogenase (BDH). A clone was selected that contained a 1.2-kb insert. The insert placed in an expression plasmid was utilized to transform Escherichia coli. These cells were shown to possess phosphatidylcholine-dependent BDH activity. Cells transformed with only the plasmid had no detectable BDH activity in the presence of phosphatidylcholine. The expressed activity in E. coli could be inhibited in a dose-dependent manner by BDH antiserum.
authors.affiliation: Department of Biology, University of Alabama, Tuscaloosa 35487-0344.
authors.forename: S
authors.initials: S
authors.lastname: Churchill
authors.forename: P
authors.initials: P
authors.lastname: Churchill
mesh_terms: D000818:Animals; D015153:Blotting, Western; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D006884:Hydroxybutyrate Dehydrogenase / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D008099:Liver / Q000201:enzymology* / Q000378:metabolism; D010713:Phosphatidylcholines / Q000494:pharmacology; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics*; D014162:Transfection; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D010713:Phosphatidylcholines; D011993:Recombinant Fusion Proteins; D006884:Hydroxybutyrate Dehydrogenase; D001616:beta-Galactosidase