• 主题
高级搜索  >
历史搜索 清空历史
首页> 美国卫生研究院文献>Scientific Reports >A genetic system for targeted mutations to disrupt and restore genes in the obligate bacterium Ehrlichia chaffeensis
【2h】

A genetic system for targeted mutations to disrupt and restore genes in the obligate bacterium Ehrlichia chaffeensis

机译：一个用于突变的遗传系统以破坏和恢复专性细菌恰菲埃里希氏体中的基因

代理获取
本网站仅为用户提供外文OA文献查询和代理获取服务，本网站没有原文。下单后我们将采用程序或人工为您竭诚获取高质量的原文，但由于OA文献来源多样且变更频繁，仍可能出现获取不到、文献不完整或与标题不符等情况，如果获取不到我们将提供退款服务。请知悉。
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Obligate intracellular bacteria (obligates) belonging to Rickettsiales and Chlamydiales cause diseases in hundreds of millions of people worldwide and in many animal species. Lack of an efficient system for targeted mutagenesis in obligates remains a major impediment in understanding microbial pathogenesis. Challenges in creating targeted mutations may be attributed to essential nature of majority of the genes and intracellular replication dependence. Despite success in generating random mutations, a method that works well in creating mutations in specific genes of interest followed by complementation remains problematic for obligates and is a highly sought-after goal. We describe protocols to generate stable targeted mutations by allelic exchange in Ehrlichia chaffeensis, an obligate intracellular tick-borne bacterium responsible for human monocytic ehrlichiosis. Targeted mutations in E. chaffeensis were created to disrupt two genes, and also to restore one gene by another allelic exchange mutation leading to the restoration of transcription and protein expression from the inactivated gene and the recovered organisms also express mCherry, which distinguishes from the wild type. We expect that the methods developed are broadly applicable to other obligates, particularly to rickettsial pathogens, to routinely perform targeted mutations to enable studies focused on protein structure-function analyses, host-pathogen interactions and in developing vaccines.
机译：属于立克次氏体和衣原体的专性细胞内细菌（专性）在全世界成千上万人和许多动物物种中引起疾病​​。缺乏专性目标诱变的有效系统仍然是理解微生物发病机理的主要障碍。产生靶向突变的挑战​​可能归因于大多数基因的本质和细胞内复制的依赖性。尽管成功地产生了随机突变，但是对于在专心的特定基因中产生突变然后进行互补的方法，效果很好，这对专性人群仍然是有问题的，并且是一个非常受欢迎的目标。我们描述协议通过等位基因交换在埃菲尔铁锈，专职负责人单核细胞埃希氏菌病的细胞内壁虱携带细菌的等位基因交换产生协议。在恰菲大肠杆菌中产生了靶向突变，以破坏两个基因，并通过另一个等位基因交换突变恢复一个基因，从而导致该基因从失活基因中恢复转录和蛋白质表达，回收的生物体也表达mCherry，这与野生动物不同类型。我们希望开发的方法广泛适用于其他专长，特别是用于立克次氏体病原体，以常规进行靶向突变，从而使研究能够专注于蛋白质结构功能分析，宿主-病原体相互作用以及开发疫苗。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
代理获取

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