Cloning and Characterization of the Bovine Gene for Steroid 21-Hydroxylase (P-450c21)

Bon Chu Chung, Karla J. Matteson, Walter L. Miller

Research output: Contribution to journalArticle

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Abstract

Steroid 21-hydroxylase activity is required for the synthesis of both cortisol and aldosterone. Using two manually synthesized oligonucleotide probes, we screened a bovine genomic DNA library and identified a phage, λE11, carrying the gene for the steroid 21-hydroxylase, P-450C21. The identity of λE11 was initially proven by initiating dideoxy sequencing from the two oligonucleotides directly on the full-length, uncleaved phage template. Hybridization of total bovine genomic DNA to λE11 restriction fragments indicates the presence of repetitive sequences in or near the P-450C21 gene. Northern blots indicate that the mature mRNA exists in two principal forms of about 2.2 and 2.4 kb in length. Southern blots indicate there are two copies of the gene in the bovine genome. Sequence analysis of a 1141-bp Eco RI fragment of the gene shows three complete exons and a portion of a fourth exon, correctly determining the amino acid sequence of 148 amino acids of this important enzyme. This cloned 1141-bp fragment cross-hybridizes to human genomic DNA, indicating it should be a useful probe for studying the human P-450C21 gene in patients having impaired 21-hydroxylase activity.

Original languageEnglish
Pages (from-to)211-219
Number of pages9
JournalDNA
Volume4
Issue number3
DOIs
Publication statusPublished - Jan 1 1985

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Steroid 21-Hydroxylase
Cloning
Organism Cloning
Genes
Bacteriophages
Exons
Deoxyribonuclease EcoRI
Genomic Library
Oligonucleotide Probes
Nucleic Acid Repetitive Sequences
DNA
Amino Acids
Southern Blotting
Aldosterone
Gene Library
Oligonucleotides
Northern Blotting
Sequence Analysis
Hydrocortisone
Amino Acid Sequence

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Genetics

Cite this

Cloning and Characterization of the Bovine Gene for Steroid 21-Hydroxylase (P-450c21). / Chung, Bon Chu; Matteson, Karla J.; Miller, Walter L.

In: DNA, Vol. 4, No. 3, 01.01.1985, p. 211-219.

Research output: Contribution to journalArticle

Chung, Bon Chu ; Matteson, Karla J. ; Miller, Walter L. / Cloning and Characterization of the Bovine Gene for Steroid 21-Hydroxylase (P-450c21). In: DNA. 1985 ; Vol. 4, No. 3. pp. 211-219.
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