Method for identifying drug-sensitizing antisense DNA fragments and use thereof

The invention provides a method for generating and selecting drug-sensitizing antisense DNA fragments. In one embodiment, the method includes identifying a gene of interest using knowledge of bacterial physiology, biochemistry, genetics, genomics, and other means. The method includes PCR amplificati...

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Bibliographische Detailangaben
Hauptverfasser: GC KEDAR, HILGERS MARK, FINN JOHN M, SHAW KAREN, BROWN-DRIVER VICKIE, STIDHAM MARK, HASELBECK ROBERT
Format: Patent
Sprache:eng
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Beschreibung
Zusammenfassung:The invention provides a method for generating and selecting drug-sensitizing antisense DNA fragments. In one embodiment, the method includes identifying a gene of interest using knowledge of bacterial physiology, biochemistry, genetics, genomics, and other means. The method includes PCR amplification of a gene of interest using genomic DNA as a template; fragmentation of the DNA by sonication or other means; selecting DNA fragments no longer than 400 base pairs; ligating the DNA fragments into a suitable expression plasmid with a selectable marker; transforming the plasmids containing the DNA fragments into the organism from which the gene of interest originated; and selecting clones from transformed cells that show a phenotypic difference of the clone grown in the presence of the inducer relative to the phenotype in the absence of inducer.