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.
MeSH terms
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Animals
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Blotting, Western
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Cloning, Molecular*
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Escherichia coli / enzymology
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Escherichia coli / genetics*
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Hydroxybutyrate Dehydrogenase / biosynthesis
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Hydroxybutyrate Dehydrogenase / genetics*
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Hydroxybutyrate Dehydrogenase / metabolism
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Liver / enzymology*
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Liver / metabolism
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Phosphatidylcholines / pharmacology
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Rats
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Recombinant Fusion Proteins / biosynthesis*
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Recombinant Fusion Proteins / genetics*
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Transfection
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beta-Galactosidase / biosynthesis
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beta-Galactosidase / genetics*
Substances
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Phosphatidylcholines
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Recombinant Fusion Proteins
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Hydroxybutyrate Dehydrogenase
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beta-Galactosidase