Molecular cloning of DNA complementary to bovine adrenal P450scc mRNA |
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Authors: | Karla J. Matteson Bon-chu Chung Walter L. Miller |
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Affiliation: | 2. Metabolic Research Unit, University of California, San Francisco, 94143 USA |
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Abstract: | P450scc is the rate-limiting hormonally regulated enzyme that cleaves the cholesterol side chain. Translation of bovine adrenocortical mRNA and immunoprecipitation with rabbit anti-bovine P450scc indicates P450scc mRNA represents 1% of the total. DNA complementary to bovine adrenocortical mRNA was cloned in the site of pBR322 by dC·dG tailing and high-efficiency transformation. A clone containing sequences complementary to P450scc mRNA was identified by hybrid-selected translation only when plasmid DNA was first purified by CsCl gradient centrifugation. As is often the case with hybrid-selected translation, the clone identified contains a small insert. |
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Keywords: | ACTH adrenocorticotrophic hormone bp base pairs kb kilobases dC deoxycytidine dG deoxyguanosine SDS sodium dodecyl sulfate |
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