Cloning of cDNA encoding steroid 11 beta-hydroxylase (P450c11).

Streamson C. Chua, Jr., P. Szabo, A. Vitek, K. H. Grzeschik, M. John, P. C. White

Research output: Contribution to journalArticle

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Abstract

We have isolated bovine and human adrenal cDNA clones encoding the adrenal cytochrome P-450 specific for 11 beta-hydroxylation (P450c11). A bovine adrenal cDNA library constructed in the bacteriophage lambda vector gt10 was probed with a previously isolated cDNA clone corresponding to part of the 3' untranslated region of the 4.2-kilobase (kb) mRNA encoding P450c11. Several clones with 3.2-kb cDNA inserts were isolated. Sequence analysis showed that they overlapped the original probe by 300 base pairs (bp). Combined cDNA and RNA sequence data demonstrated a continuous open reading frame of 1509 bases. P450c11 is predicted to contain 479 amino acid residues in the mature protein in addition to a 24-residue amino-terminal mitochondrial signal sequence. A bovine clone was used to isolate a homologous clone with a 3.5-kb insert from a human adrenal cDNA library. A region of 1100 bp was 81% homologous to 769 bp of the coding sequence of the bovine cDNA except for a 400-bp segment presumed to be an unprocessed intron. Hybridization of the human cDNA to DNA from a panel of human-rodent somatic cell hybrid lines and in situ hybridization to metaphase spreads of human chromosomes localized the gene to the middle of the long arm of chromosome 8. These data should be useful in developing reagents for heterozygote detection and prenatal diagnosis of 11 beta-hydroxylase deficiency, the second most frequent cause of congenital adrenal hyperplasia.

Original languageEnglish (US)
Pages (from-to)7193-7197
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume84
Issue number20
StatePublished - Oct 1987
Externally publishedYes

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Steroid 11-beta-Hydroxylase
Organism Cloning
Complementary DNA
Clone Cells
Base Pairing
Gene Library
Heterozygote Detection
Bacteriophage lambda
Congenital Adrenal Hyperplasia
Chromosomes, Human, Pair 8
Hybrid Cells
Human Chromosomes
3' Untranslated Regions
Hydroxylation
Metaphase
Protein Sorting Signals
Prenatal Diagnosis
Introns
Open Reading Frames
In Situ Hybridization

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

Cloning of cDNA encoding steroid 11 beta-hydroxylase (P450c11). / Chua, Jr., Streamson C.; Szabo, P.; Vitek, A.; Grzeschik, K. H.; John, M.; White, P. C.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 84, No. 20, 10.1987, p. 7193-7197.

Research output: Contribution to journalArticle

Chua, Jr., Streamson C. ; Szabo, P. ; Vitek, A. ; Grzeschik, K. H. ; John, M. ; White, P. C. / Cloning of cDNA encoding steroid 11 beta-hydroxylase (P450c11). In: Proceedings of the National Academy of Sciences of the United States of America. 1987 ; Vol. 84, No. 20. pp. 7193-7197.
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