日本人ブル-ム症候群の分子的欠陥

日本布卢姆综合征的分子缺陷

• 批准号: 03151031
• 负责人: 巽 紘一
• 金额: $ 4.99万
• 依托单位: Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Cancer Research
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03151031/
• 关键词: ブル-ム症候群; リンパ芽球様細胞; DNAリガ-ゼI; 6チオグアニン抵抗性; HPRT; 欠失; ポリ(ADP.リボ-ス)シンセタ-ゼ; RFLP

ブル-ム症候群(Bloom's syndrome BS)患者および正常人由来のリンパ芽球様細胞(LCL)の粗抽出液中のDNAリガ-ゼI活性をHAカラムクロマトグラフィ-によりリガ-ゼII活性と分離して測定した。正常人由来7株、日本人BS由来5株、白人BS由来1株、ユダヤ人BS由来3株のリガ-ゼI活性は標準NLーHa株活性の比で示すと正常人0.74ー1.24平均0.91、BS0.33ー1.12平均0.65と幅広く分布した。BSF07(日本人)は明らかに正常人より低値を示すが、他のBS株では正常人の活性範囲を有意に外れるものはなく、また活性値の高低に民族差を認めなかった。リガ-ゼI反応の基質オリゴdTおよびATPに対するKm値でも正常人とBS間に差はなかった。ヒトDNAリガ-ゼIcDNAを用いてBS7株と正常人株のRFLP(EcoRI,BamHI,XbaI)検索を行ったが差異を認めなかった。DNAリガ-ゼIに関する数個のゲノム・クロ-ンを得たが5'上流の調節領域を含んでいなかった。6チオグアニン抵抗性(TG)変異を指標とした突然変異率測定でBS2KA 19.4x10^<-7>、GM3403 13.9x10^<-7>/細胞/世代と正常人細胞の5〜10倍高値を示し、multiplex PCRによるHPRT各exonの構造解析から、調べた8個のTGクロ-ンすべてに欠失を認めた。この内約半数が全exonの欠失であった。BS患者株粗抽出液中のポリ(ADP・リボ-ス)シンセタ-ゼ活性は対照正常人細胞株との間に有意差はなかった。

Isolation and determination of DNA activity in crude extracts from human embryonic cells (LCL) from patients with Bloom's syndrome BS and normal individuals 7 strains from normal individuals, 5 strains from Japanese BS, 1 strain from white BS, 3 strains from Japanese BS, and the ratio of activity to standard NL strains was 0.74 to 1.24, averaging 0.91, and BS to 0.33 to 1.12, averaging 0.65. BSF07(Japanese) is a normal person with low activity, and his BS strain is a normal person with high activity and low activity. The difference between normal people and BS is the difference between normal people and ATP. The difference between the DNA fragment of BS7 strain and that of normal human strain was identified by RFLP(EcoRI,BamHI,XbaI). The DNA sequence has several regulatory domains, including the 5'upstream regulatory domain. 6. The variation index of resistance (TG) was determined by the sudden variation rate of BS2KA 19.4x10^<-7>, GM3403 13.9x10^<-7>/cell/generation and 5 ~ 10 times higher than that of normal human cells. About half of them are missing. The activity of ADP in crude extracts of BS patients was significantly different from that of normal human cells.

项目成果

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