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請使用永久網址來引用或連結此文件:
http://ir.cmu.edu.tw/ir/handle/310903500/28729
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| 題名: | Identification of the copper-zinc superoxide dismutase activity in Mycoplasma hyopneumoniae |
| 作者: | Chen, JR;Weng, CN;Ho, TY;Cheng, IC;Lai, SS |
| 貢獻者: | 中醫學院中醫所;Natl Taiwan Univ, Dept Vet Med, Taipei 106, Taiwan;Natl Taiwan Univ, Pig Res Inst, Dept Pathobiol, Taipei 106, Taiwan;China Med Coll, Inst Chinese Med Sci, Taichung, Taiwan |
| 日期: | 2000 |
| 上傳時間: | 2010-09-24 13:44:16 (UTC+8) |
| 出版者: | ELSEVIER SCIENCE BV |
| 摘要: | Background: A single nonfunctional chimeric gene with its 5' and 3' ends corresponding to CYP21P and CYP21, respectively, is caused by unequal gene crossover in the CYP21 genes during meiosis. The presence of the chimeric CYP21P/CYP21 molecule can not be detected by conventional FCR methods and therefore may be lost in PCR amplification. This leads to a false result and diagnostic discordance. Methods: We developed a rapid and direct method to detect a chimeric CMP21P/CYP21 gene that uses a 3'-specific primer for the CYP21 gene and two different 5' primers for both CYP21 and CYP21P to amplify the wild-type CYP21 and the chimeric CYP21P/CYP21 genes. A secondary PCR that can differentiate the chimeric from the wild-type gene was also performed. The PCR product was directly analyzed on agarose gel. Results: After careful titration, we found that earlier failure to detect the chimeric CYP21P/CYP21 gene could be caused by unequal concentrations of two independent alleles as the PCR template or by the lack of primers to amplify chimeric molecules. We successfully amplified the chimeric gene using our improved method. Conclusions: The chimeric CYP21P/CYP21 is present in a large portion of congenital adrenal hyperplasia patients. By adding a CYP21P/CYP21-specific primer, we were able to amplify and detect both homozygous and heterozygous chimeric genes. Therefore, our new PCR-based assay is a more effective way to analyze congenital adrenal hyperplasia mutations. (C) 2000 American Association for Clinical Chemistry. |
| 關聯: | VETERINARY MICROBIOLOGY 73(4):301-310 |
| 顯示於類別: | [中國醫學研究所] 期刊論文
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