• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Genomics >Integrity of human YACs during propagation in recombination-deficient yeast strains.
【24h】

Integrity of human YACs during propagation in recombination-deficient yeast strains.

机译：人YAC在重组缺陷型酵母菌株中繁殖期间的完整性。

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Several isogenic strains with defects in recombination/repair genes (RAD1, RAD50, RAD51, RAD52, RAD54, and RAD55) were examined for their ability to propagate accurately a variety of linear and circular yeast artificial chromosomes (YACs) containing human DNA inserts. To assess YAC stability, the human DNA inserts were internally marked by an ADE2-pBR-URA3 cassette. Following selection for Ura- clones on 5-fluoroorotic acid containing medium, the following types of YAC deletions were identified: (i) those caused by homologous recombination with a telomeric pBR sequence; (ii) internal deletions, presumed to occur by recombination between commonly occurring DNA repeats such as Alu and LINE sequences; and (iii) deletions leading to loss of part of a YAC arm. rad52 host strains, but not other recombination-deficient strains, decreased the rate of all types of YAC deletions 25- to 400-fold. We have also developed and tested kar1 strains with a conditional RAD52 gene that allow transfer of a YAC from any host into a recombination-deficient background. These strains provide an efficient tool for stabilization of YACs and are useful for allowing additional recombinational modification of YACs. Copyright 1999 Academic Press.
机译：检查了几个在重组/修复基因中存在缺陷的同基因菌株（RAD1，RAD50，RAD51，RAD52，RAD54和RAD55）的能力，使其能够准确繁殖各种包含人类DNA插入片段的线性和环状酵母人工染色体（YAC）。为了评估YAC的稳定性，人类DNA插入片段内部用ADE2-pBR-URA3盒进行了标记。在含有5-氟乳清酸的培养基上选择Ura-克隆后，鉴定出以下类型的YAC缺失：（i）与端粒pBR序列同源重组引起的YAC缺失; （ii）内部缺失，推测是由于常见的DNA重复序列如Alu和LINE序列之间的重组而发生的; （iii）导致YAC臂部分丢失的删除。 rad52宿主菌株，而不是其他缺乏重组的菌株，将所有类型的YAC缺失率降低了25到400倍。我们还开发并测试了带有条件RAD52基因的kar1菌株，该基因可将YAC从任何宿主转移到重组缺陷的背景中。这些菌株为稳定YAC提供了有效的工具，可用于允许YAC进行其他重组修饰。版权所有1999 Academic Press。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号