Genetic footprinting with mariner-based transposition in Pseudomonas aeruginosa
AUTOR(ES)
Wong, Sandy M.
FONTE
The National Academy of Sciences
RESUMO
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-SceI 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. aeruginosa, validating the practicality of this comparative genomics strategy to identify essential genes in P. aeruginosa.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=27802Documentos Relacionados
- In vivo transposition of mariner-based elements in enteric bacteria and mycobacteria
- Development of a mariner-Based Transposon for Use in Sorangium cellulosum
- Transposition of the mariner element from Drosophila mauritiana in zebrafish
- Genetic heterogeneity in strains of Pseudomonas aeruginosa from patients with cystic fibrosis.
- FERTILITY FACTORS AND GENETIC LINKAGE IN PSEUDOMONAS AERUGINOSA