IQUIBICEN 23947
INSTITUTO DE QUIMICA BIOLOGICA DE LA FACULTAD DE CIENCIAS EXACTAS Y NATURALES
Unidad Ejecutora - UE
artículos
Título:
Genetic Manipulation of Lytic Bacteriophages with BRED: Bacteriophage Recombineering of Electroporated DNA
Autor/es:
PIURI, MARIANA; HATFULL, GRAHAM F.; MARINELLI, LAURA J.
Revista:
METHODS IN MOLECULAR BIOLOGY (CLIFTON, N.J.)
Editorial:
Humana Press Inc.
Referencias:
Año: 2019 vol. 1898 p. 69 - 80
ISSN:
1064-3745
Resumen:
We describe a recombineering-based method for the genetic manipulation of lytically replicating bacteriophages, focusing on mycobacteriophages. The approach utilizes recombineering-proficient strains of Mycobacterium smegmatis and employs a cotransformation strategy with purified phage genomic DNA and a mutagenic substrate, which selects for only those cells that are competent to take up DNA. The cotransformation method, combined with the high rates of recombination obtained in M. smegmatis recombineering strains, allows for the efficient and rapid generation of bacteriophage mutants.
