Structure and expression of the CYP21 (p450c21, steroid 21-hydroxylase) gene with respect to its deficiency

Bon Chu Chung, Meng Chun Hu, Victor M. Guzov, Du An Wu

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Steroid 21-hydroxylase (P450c21) deficiency is the major cause of a common genetic disease, congenital adrenal hyperplasia, with the symptoms of virilization due to steroid imbalance. We have devised a fast diagnostic method to detect common mutations in the c21B gene by a two-step gene amplification procedure coupled to restriction digestion. This procedure does not require isotopes and is suitable for routine use in a hospital setting. In addition, we have developed a procedure for the production of active P450c21 in E. coli. We tested many different vector and bacterial strain combinations to find out the best condition for P450c21 expression. The bacteria harboring the P450c21 expression plasmid were grown in a rich media supplemented with trace metals, heme biosynthesis precursor δlevulinic acid, and induced with IPTG at 20°C for 48 h. We found that low growth temperature and long induction time were important for abundant synthesis of P450c21 in E. coli.

Original languageEnglish
Pages (from-to)343-352
Number of pages10
JournalEndocrine Research
Volume21
Issue number1-2
DOIs
Publication statusPublished - Jan 1 1995

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Steroid 21-Hydroxylase
Genes
Escherichia coli
Virilism
Isopropyl Thiogalactoside
Congenital Adrenal Hyperplasia
Inborn Genetic Diseases
Gene Amplification
Heme
Isotopes
Digestion
Plasmids
Metals
Steroids
Bacteria
Mutation
Temperature
Growth

All Science Journal Classification (ASJC) codes

  • Endocrinology

Cite this

Structure and expression of the CYP21 (p450c21, steroid 21-hydroxylase) gene with respect to its deficiency. / Chung, Bon Chu; Hu, Meng Chun; Guzov, Victor M.; Wu, Du An.

In: Endocrine Research, Vol. 21, No. 1-2, 01.01.1995, p. 343-352.

Research output: Contribution to journalArticle

Chung, Bon Chu ; Hu, Meng Chun ; Guzov, Victor M. ; Wu, Du An. / Structure and expression of the CYP21 (p450c21, steroid 21-hydroxylase) gene with respect to its deficiency. In: Endocrine Research. 1995 ; Vol. 21, No. 1-2. pp. 343-352.
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