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Titolo:
Genetic footprinting with mariner-based transposition in Pseudomonas aeruginosa
Autore:
Wong, SM; Mekalanos, JJ;
Indirizzi:
Harvard Univ, Sch Med, Dept Microbiol & Mol Genet, Boston, MA 02115 USA Harvard Univ Boston MA USA 02115 robiol & Mol Genet, Boston, MA 02115 USA Harvard Univ, Sch Med, Shipley Inst Med, Boston, MA 02115 USA Harvard Univ Boston MA USA 02115 , Shipley Inst Med, Boston, MA 02115 USA
Titolo Testata:
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
fascicolo: 18, volume: 97, anno: 2000,
pagine: 10191 - 10196
SICI:
0027-8424(20000829)97:18<10191:GFWMTI>2.0.ZU;2-9
Fonte:
ISI
Lingua:
ENG
Soggetto:
GRAM-NEGATIVE BACTERIA; DOUBLE-STRAND BREAKS; GROUP-I INTRON; HOMOLOGOUS RECOMBINATION; ESCHERICHIA-COLI; CYSTIC-FIBROSIS; ENDONUCLEASE; MUTAGENESIS; VECTORS; YEAST;
Keywords:
transposon; SCE jumping;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
31
Recensione:
Indirizzi per estratti:
Indirizzo: Mekalanos, JJ Harvard Univ, Sch Med, Dept Microbiol & Mol Genet, 200 Longwood Ave, Boston, MA 02115 USA Harvard Univ 200 Longwood Ave Boston MA USA 02115 02115 USA
Citazione:
S.M. Wong e J.J. Mekalanos, "Genetic footprinting with mariner-based transposition in Pseudomonas aeruginosa", P NAS US, 97(18), 2000, pp. 10191-10196

Abstract

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 toidentify 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 andefficient allelic exchange system by using the I-Scel homing endonuclease in conjunction with in vitro mariner mutagenesis to generate mutants withintargeted regions of the P. aeruginosa chromosome for genetic footprinting analyses. This technique for generating transposon insertion mutants shouldbe widely applicable to other organisms that are not naturally transformable or may lack well developed in vivo transposition systems. We tested thissystem 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.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 28/03/20 alle ore 23:39:44