A rapid and simple method for inactivating chromosomal genes in Yersinia.
FEMS Immunol Med Microbiol
; 38(2): 113-6, 2003 Sep 22.
Article
em En
| MEDLINE
| ID: mdl-13129645
ABSTRACT
A polymerase chain reaction (PCR)-based procedure without any cloning step was developed for a rapid mutagenesis/deletion of chromosomal target genes in Yersinia. For this purpose, a PCR fragment carrying an antibiotic resistance gene flanked by regions homologous to the target locus is electroporated into a recipient strain expressing the highly proficient homologous recombination system encoded by plasmid pKOBEG-sacB. Two PCR procedures were tested to generate an amplification product formed of an antibiotic resistance gene flanked by short (55 bp) or long (500 bp) homology extensions. Using this method, three chromosomal loci were successfully disrupted in Yersinia pseudotuberculosis. The use of this technique allows rapid and efficient large-scale mutagenesis of Yersinia target chromosomal genes.
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Bases de dados:
MEDLINE
Assunto principal:
Proteínas de Bactérias
/
Yersinia pseudotuberculosis
/
Resistência a Canamicina
/
Reação em Cadeia da Polimerase
/
Mutagênese
/
Deleção de Genes
Tipo de estudo:
Evaluation_studies
Idioma:
En
Revista:
FEMS Immunol Med Microbiol
Assunto da revista:
ALERGIA E IMUNOLOGIA
/
DOENCAS TRANSMISSIVEIS
/
MICROBIOLOGIA
Ano de publicação:
2003
Tipo de documento:
Article
País de afiliação:
França