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Deletion of the C4-CYP21 Repeat Module Leading to the Formation of a Chimeric CYP21P/CYP21 Gene in a 9.3-kb Fragment as a Cause of Steroid 21-Hydroxylase Deficiency

¹ King Car Food Industrial Co., Ltd., Yuan-Shan Research Institute, No. 326, Yuan Shan Rd., Sec. 2, Yuan Shan, Ilan 264, Taiwan, Republic of China;
² Graduate Institute of Cell and Molecular Biology, Taipei Medical University, Taipei 110, Taiwan, Republic of China;
³ Department of Pediatrics, Mackay Memorial Hospital, Taipei 104, and College of Medicine, Taipei Medical University, Taipei 110, Taiwan, Republic of China;
⁴ Genetika-Centro de Aconselhamento e Laboratorio de Genetica, Curitiba, Parana, Brazil 80410;
⁵ Department of Pediatrics, College of Medicine, National Cheng Kung University, Tainan 704, Taiwan, Republic of China;
⁶ Department of Pediatrics, Division of Genetics, Kaohsiung Medical University Hospital, Kaohsiung 807, Taiwan, Republic of China;
⁷ Division of Endocrinology, Department of Pediatrics, Chang Gung Children’s Hospital, Taoyuan 330, Taiwan, Republic of China;

^aauthor for correspondence: fax 886-3-9228030, e-mail hhlee@ms2.kingcar.com.tw

The first 300 words of the full text of this article appear below.

Gross gene deletions have been reported in 20% of alleles in patients with congenital adrenal hyperplasia (CAH) involving a 21-hydroxylase deficiency (1). This type of deletion occurs in the RCCX module, including the CYP21P, tenascin A (TNXA), RP2, C4B, CYP21, and tenascin B (TNXB) genes, as evidenced by a 30-kb deletion identified by pulse-field electrophoresis (2). Inactivation of the CYP21 gene may also occur through intergenic recombination with transfer of deleterious mutations from the neighboring CYP21P pseudogene. The frequency of gene deletions or conversions in CAH is controversial (3)(4)(5) and is dependent on the population studied. Evidence for gene deletions and/or conversions is traditionally obtained by Southern blot analysis. Multiple probes and separate restriction endonuclease digestions are used. TaqI generates 3.7-kb (functional) and 3.2-kb (pseudogene) fragments, and BglII produces 11-kb (functional) and 12-kb (pseudogene) fragments. These analyses have been used since 1984 (1)(3)(5)(6)(7)(8)(9). However, the method is indirect and time-consuming, and densitometry of fragments can be prone to error.

To identify the interchange region and improve detection of gene deletions and conversions in the RCCX module (10)(11)(12)(13), we have developed a novel Southern blot analysis that uses two restriction endonucleases, AseI and NdeI, and requires only one probe. In addition, we use a PCR product amplified with locus-specific primers covering the TNXB gene to the 5' end of CYP21P or CYP21 to directly analyze the 3.2/3.7-kb TaqI fragment and the status of the CYP21 gene.

For the novel Southern blot analysis, 10 µg of genomic DNA was digested with the restriction endonucleases AseI and . . . [Full Text of this Article]

The following articles in journals at HighWire Press have cited this article:

				S. Parajes, C. Quinterio, F. Dominguez, and L. Loidi A Simple and Robust Quantitative PCR Assay to Determine CYP21A2 Gene Dose in the Diagnosis of 21-Hydroxylase Deficiency Clin. Chem., September 1, 2007; 53(9): 1577 - 1584. [Abstract] [Full Text] [PDF]

				H.-H. Lee, Y.-J. Lee, P. Chan, and C.-Y. Lin Use of PCR-Based Amplification Analysis as a Substitute for the Southern Blot Method for CYP21 Deletion Detection in Congenital Adrenal Hyperplasia Clin. Chem., June 1, 2004; 50(6): 1074 - 1076. [Full Text] [PDF]

				P. F.J. Koppens, H. J. Degenhart, and H.-H. Lee PCR-based Detection of CYP21 Deletions * One of the authors of the Technical Brief cited above responds: Clin. Chem., September 1, 2003; 49(9): 1555 - 1557. [Full Text] [PDF]

				H.-H. Lee, S.-F. Chang, F.-J. Tsai, L.-P. Tsai, and C.-Y. Lin Mutation of IVS2 -12A/C>G in Combination with 707-714delGAGACTAC in the CYP21 Gene Is Caused by Deletion of the C4-CYP21 Repeat Module with Steroid 21-Hydroxylase Deficiency J. Clin. Endocrinol. Metab., June 1, 2003; 88(6): 2726 - 2729. [Abstract] [Full Text] [PDF]