Abstract
21 hydroxylase is a single specific 49000 dalton microsomal cytochrome P450 termed P450c21. Using two chemically synthesized oligonucleotide probes, we screened a bovine genomic DNA library, and identified a phage, λE11, carrying the gene for P450c21. The identity of λE11 was 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 some DNA in or near the P450c21 gene contains repetitive sequences. Northern blots indicate the primary gene transcript is 6.8 to 9.5 kb long and 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 EcoRI fragment of the gene shows three complete exons and a portion of a fourth exon. The intron/exon arrangement is similar to that seen for the P450c gene induced in liver by 3-methylcholanthrene but not to that of the P450e gene induced by phenobarbital. This sequence analysis correctly determines 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 is a useful probe for studying the human P450c21 gene in patients having 21 hydroxylase deficiency.
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Chung, BC., Matteson, K. & Miller, W. 119 CLONING AND STRUCTURE OF THE BOVINE 21 HYDROXYLASE GENE. Pediatr Res 19, 623 (1985). https://doi.org/10.1203/00006450-198506000-00139
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DOI: https://doi.org/10.1203/00006450-198506000-00139