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目的 研究 46,XY女性性反转患者发病的分子机理。方法 应用聚合酶链反应 (polymerasechain reaction,PCR)扩增 1例性反转患者和其父亲的 SRY基因片段 ;PCR产物连接到 p UCm-T载体上 ,在 ABI 3 77-3自动测序仪上完成测序以查明突变 ;应用 PCR-限制性酶切对 DNA测序的结果进行检测。结果 发现该患者的 SRY基因存在点突变 (T3 87A) ,导致 SRY蛋白发生氨基酸翻译终止 (酪氨酸 12 9终止密码 ) ,患者父亲的序列正常。由于患者 SRY序列中增加一个 Mae 酶切位点 ,PCR-限制性内切酶酶切电泳检测 ,结果显示患者出现 3条带 (13 1bp、2 3 1bp和 2 47bp) ,而正常人出现 2条带 (13 1bp和 478bp) ,进一步验证了序列分析的结果。经查证数据库 ,该突变是一个未见报道的新型 SRY基因突变。结论 这一新型突变的发现 ,有助于进一步阐明 46,XY女性性反转发病的分子机理
Objective To study the molecular mechanism of the pathogenesis of XY female sexual inversion patients. Methods A SRY gene fragment was amplified from a patient and its father by polymerase chain reaction (PCR). The PCR product was ligated into pUCm-T vector and sequenced on an ABI 3 77-3 automated sequencer Sequencing was performed to identify the mutations; PCR sequencing was used to detect DNA sequencing results. The results showed that there was a point mutation in the SRY gene (T3 87A), resulting in the termination of amino acid translation of the SRY protein (tyrosine 129 stop codon). The patient’s father’s sequence was normal. As a result of the addition of a Mae restriction site in the SRY sequence of the patient, PCR-restriction endonuclease digestion electrophoresis revealed that there were 3 bands (13 1bp, 2 3 1bp and 2 47bp) in the patient and 2 in the normal Bands (13 lbp and 478 bp) further validated the results of the sequence analysis. After verification database, the mutation is an undiscovered new SRY gene mutation. Conclusion The discovery of this new mutation will help further elucidate the molecular mechanism of the pathogenesis of sexual inversion in women46 and XY