Random Transposon Mutagenesis for Cell-Envelope Resistant to Phage Infection.
In order to identify host components involved in the infective process of bacteriophages, we developed a wide-range strategy to obtain cell envelope mutants, using Escherichia coli W3110 and its specific phage mEp213. The strategy consisted in four steps: (1) random mutagenesis using transposon miniTn10Km(r); (2) selection of phage-resistant mutants by replica-plating; (3) electroporation of the phage-resistant mutants with mEp213 genome, followed by selection of those allowing phage development; and (4) sequencing of the transposon-disrupted genes. This strategy allowed us to distinguish the host factors related to phage development or multiplication within the cell, from those involved in phage infection at the level of the cell envelope.
Departamento de Genética y Biología Molecular, Centro de Investigación y de Estudios Avanzados del IPN (CINVESTAV-IPN), C.P., México, DF, Mexico.
Ruth
R
Reyes-Cortés
Departamento de Genética y Biología Molecular, Centro de Investigación y de Estudios Avanzados del IPN (CINVESTAV-IPN), C.P., México, DF, Mexico.
Emma S
ES
Arguijo-Hernández
Departamento de Genética y Biología Molecular, Centro de Investigación y de Estudios Avanzados del IPN (CINVESTAV-IPN), C.P., México, DF, Mexico.
Marco A
MA
Carballo-Ontiveros
Departamento de Genética y Biología Molecular, Centro de Investigación y de Estudios Avanzados del IPN (CINVESTAV-IPN), C.P., México, DF, Mexico.
Eva
E
Martínez-Peñafiel
Departamento de Genética y Biología Molecular, Centro de Investigación y de Estudios Avanzados del IPN (CINVESTAV-IPN), C.P., México, DF, Mexico. luisk@cinvestav.mx.
Luis
L
Kameyama
D001435:Bacteriophages / Q000235:genetics / Q000254:growth & development / Q000472:pathogenicity*; D002473:Cell Wall / Q000235:genetics / Q000378:metabolism; D004251:DNA Transposable Elements; D018274:Electroporation; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development* / Q000821:virology; D029968:Escherichia coli Proteins / Q000235:genetics*; D016254:Mutagenesis, Insertional; D010948:Viral Plaque Assay
D004251:DNA Transposable Elements; D029968:Escherichia coli Proteins
Bacterial receptor; Bacteriophages; Cell-envelope; Infective process; Random mutagenesis