权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Analysis of Radiation Hypersensitivity of Human Cells From Genetic Disease by the use of Chromosome Transfer

利用染色体转移分析遗传性疾病引起的人体细胞辐射超敏反应

基本信息

批准号：
01580209
负责人：
EJIMA Yosuke
金额：
$ 1.73万
依托单位：
Kyoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
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

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01580209/
关键词：
Radiosensitivity Human genetic disease Chromosome transfer

项目摘要

The chromosomal localization of the gene which complements radiation hypersensitivity of AT(ataxia telangiectasia)cells was studied by microcell-mediated chromosome transfer.1. An immortalized AT cell line, AT2KYSV, aDd its 6-thioguanineresistant derivative, AT2KYSVTG, were used as recipients. Five strains of mouse A9 cells carrying a single human chromosome were used as chromosome donors. The chromosomes carried were PSV2neo-tagged human chromosome 11, X chromosome, or X/11 recombinant chromosomes containing either llpter-ql3, llpter-q23 or llpll-qter.2. Among seven G418-resistant microcell hybrids obtained from the transfer of a PSV2neo-tagged chromosome 11, three clones showed restoration of radiation resistance with concomitant gain of an extra chromosome 11, while the others contained no recognizable or deleted chromosome 11.3. The HAT-resistant microcell hybrids obtained from the transfer of an intact X chromosome or an X/11 chromosome bearing the Ilpter-ql3 region did not show a diffprence in radiosensitivity from parental AT cells, while those obtained from the transfer of X/11 chromosomes bearing either the llpll-qter or Ilpter-q23 region exhibited a marked radioresistance.4. A HAT-resistant but radiosensitive variant was further obtained from the microcell fusion with an A9 strain carrying an X/11 chromosome bearing llpll-qter region, in which a deletion at the llq23 region was found. The results indicate that the gene which complements a radiosensitive phenotype of AT is located at the q23 region of chromosome 11. The derivative chromosome 11 found here to be associated with concomitant loss of the AT locus or its function would be useful for the molecular dissection of the llq23 region with relevance to the AT gene.

本研究采用微细胞介导的染色体转移技术，对共济失调毛细血管扩张症（ataxia telangiectasia，AT）细胞辐射超敏反应互补基因进行了染色体定位.以永生化AT细胞系AT 2KYSV及其6-硫代鸟嘌呤抗性衍生物AT 2KYSVTG为受体。将携带单个人染色体的五种小鼠A9细胞株用作染色体供体。携带的染色体是PSV 2neo标记的人11号染色体、X染色体或含有11 pter-q13、11 pter-q23或11 pll-qter的X/11重组染色体。在7个G418抗性微细胞杂种从转移的PSV 2neo标记的染色体11，三个克隆表现出恢复辐射抗性伴随额外的染色体11的增益，而其他人不包含可识别的或删除的染色体11.3。由完整的X染色体或携带Ilpter-q13区域的X/11染色体转移获得的HAT抗性微细胞杂种与亲本AT细胞的辐射敏感性没有差异，而由携带Ilpter-q23区域或IIpter-q23区域的X/11染色体转移获得的HAT抗性微细胞杂种表现出显著的辐射抗性。通过与携带有X/11染色体的A9株的微细胞融合，进一步获得了一个抗AT但对辐射敏感的突变体，该突变体在11 q23区有缺失，结果表明，与AT辐射敏感表型互补的基因位于11号染色体的q23区。这里发现的衍生染色体11与AT基因座或其功能的伴随丧失相关，这将有助于与AT基因相关的llq 23区域的分子解剖。