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ステロイドホルモン受容体異常症の分子遺伝学的解析とDNA診断

类固醇激素受体异常的分子遗传学分析和DNA诊断

基本信息

批准号：
01480292
负责人：
名和田新
金额：
$ 1.28万
依托单位：
Kyushu University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1991
项目状态：
已结题

项目摘要

私達は結合実験より睾丸性女性化症完全型ではレセプタ-欠損Reifーensteint症候群ではレセプタ-数、親和性共に正常で、熱に不安定なアンドロゲンレセプタ-異常を証明した。AR DNAの各エクソンについて、それぞれの近傍部イントロンに設定したプライマ-を用い、患者ゲノムDNAをPCRにて増幅後、塩基配列を決定した。完全型睾丸女性化症では786Met(ATG)→786Val(GTG)と、Reifenstein症候群では743Gly(GGG)→743Val(GTG)の点突然変異によるーアミノ酸置換を証明した。この点突然変異が実際アンドロゲンレセプタ-の作用発現において異常を示すかどうかを明らかにするため、siteーdeirected mutagenesisにより完全型睾丸女性化症786Val及びReifenstein症候群743Valの突然変異を持つアンドロゲンレセプタ-cDNAを作成し、これをpCMVに挿入し、COS1細胞にtransfectし合成したアンドロゲンレセプタ-を検索すると、完全型睾丸性女性化症及びReifenstein症候群共にWestern blotでは正常と同じ大きさと量のレセプタ-蛋白が証明されるが、787Valは ^3HR1881とはほとんど結合せず,743ValはARの減少と熱不安性を示す事が証明された。これらの結果は患者線維芽細胞のARの性質と一致した。この変異レセプタ-のtranscripーtion活性を明らかにするため、MMTVーCATと変異レセプタ-遺伝子pCMVMーTARH(Val786,又はVal443)をCOS1細胞にcontransfectし、CAT測定をした。DHT添加により正常(WT)ではCAT活性が著明に発現しているが,完全型睾丸性女性化症(TF)では全く見られず、Reifenstein症候群(RF)ではWTに比べると弱いが若干CAT活性が見られる。これらの結果はAR遺伝子の点突然変異と結合実験の結果と、その発現型との関係を良く説明する興味ある結果を得た。

The combination of testicular feminization complete form, partial impairment Reif ensteint syndrome, partial number and affinity is normal, and thermal instability is abnormal. AR DNA is used to determine the sequence of DNA fragments, amplification and nucleotide sequence of AR DNA fragments. Complete testicular feminization is demonstrated by the abrupt change of 786 Met (ATG) → 786 Val (GTG) → Reifenstein syndrome 743 Gly (GGG) → 743 Val (GTG) →The sudden change of site in complete testicular feminization 786 Val and Reifenstein syndrome 743 Val is caused by the sudden change of site in complete testicular feminization 786 Val and Reifenstein syndrome 743 Val, and the sudden change of site in complete testicular feminization 786 Val and Reifenstein syndrome 743 Val is caused by the sudden change of site in complete testicular feminization 786 Val and Reifenstein syndrome 743 Val. The sudden change of site in complete testicular feminization 786 Val is caused by the sudden change of site in complete testicular feminization 786 Val and Reifenstein syndrome 743 Val. The sudden change of site in complete testicular feminization 786 Val and Reifenstein syndrome 743 Val is caused by the sudden change of site in complete testicular feminization 786 Val and Reifenstein syndrome 743 Val. The sudden change of site in complete testicular feminization 786 Val is caused by the sudden change of site in complete testicular feminization 786 Val and Reifenstein syndrome 743 Val. The sudden change of site in complete testicular feminization 786 Val is caused by Complete testicular feminization and Reifenstein syndrome were demonstrated by Western blot analysis of normal, high, and low levels of protein, 787 Val, 3HR1881, and 743 Val, respectively. The results showed that the AR properties of the cells in the patient's lineal cells were consistent. The transfect-specific activity of these proteins was measured in COS-1 cells by MMTV CAT and pCMVM TARH (Val 786, Val 443). CAT activity was significantly higher in normal (WT), complete testicular feminization (TF) and Reifenstein syndrome (RF) than in WT. The result of this study is that the AR gene is suddenly different from the result of the study, and the relationship between the two is well explained.