Patent ID: 7709196

Claim:
A method for identifying functional mutant ribosomes that may be suitable as drug targets comprising: (a) transforming a first set of host cells with a first set of plasmids, each plasmid comprising a first mutant rRNA gene and a first selectable marker gene; wherein said first mutant rRNA gene comprises at least one mutation in addition to a first mutant ribosome binding sequence; and said first selectable marker gene comprises a first mutant message binding sequence; and wherein said first mutant ribosome binding sequence and said first mutant message binding sequence are a mutually compatible pair; thereby forming a first set of transformed host cells; (b) isolating from the first set of transformed host cells those host cells which express the selectable marker gene product; and (c) sequencing the first mutant rRNA gene from each host cell isolated in step (b), thereby identifying functionally important target regions of interest, wherein said functionally important target regions of interest comprise nucleotide sequences of one or more nucleic acids or nucleotide motifs which are conserved in each first mutant rRNA gene sequenced; (d) generating a second plurality of mutant rRNA genes wherein the regions of interest of step (c) are mutated; and each rRNA gene further comprises a second mutant ribosome binding sequence; (e) inserting the second plurality of mutant rRNA genes comprising the mutated regions of interest from step (d) into a second plurality of plasmids; wherein said plasmids further comprise a second genetically engineered gene which encodes a second selectable marker having a second mutant message binding sequence, wherein the second mutant ribosome binding sequence and the second mutant message binding sequence are a mutually compatible pair; (f) transforming a second set of host cells with the plasmids from step (e), thereby forming a second set of transformed host cells; (g) isolating from the second set of transformed host cells of step (f) those host cells which express the selectable marker gene product; and (h) sequencing the rRNA gene from each host cell isolated in step (g), thereby identifying functional mutant ribosomes that may be suitable as drug targets.