期刊
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
卷 97, 期 18, 页码 10191-10196出版社
NATL ACAD SCIENCES
DOI: 10.1073/pnas.97.18.10191
关键词
transposon; SCE jumping
资金
- NIAID NIH HHS [R01 AI026289] Funding Source: Medline
The complete DNA sequence of Pseudomonas aeruginosa provides an opportunity to apply functional genomics to a major human pathogen, A comparative genomics approach combined with genetic footprinting was used as a strategy to identify genes required for viability in P. aeruginosa, Use of a highly efficient in vivo mariner transposition system in P. aeruginosa facilitated the analysis of candidate genes of this class. We have developed a rapid and efficient allelic exchange system by using the I-Scel homing endonuclease in conjunction with in vitro mariner mutagenesis to generate mutants within targeted regions of the P. aeruginosa chromosome for genetic footprinting analyses. This technique for generating transposon insertion mutants should be widely applicable to other organisms that are not naturally transformable or may lack well developed in vivo transposition systems. We tested this system with three genes in P, aeruginosa that have putative essential homologs in Haemophilus influenzae, We show that one of three H. influenzae essential gene homologs is needed for growth in P, aeroginosa, validating the practicality of this comparative genomics strategy to identify essential genes in P. aeruginosa.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据