LS4 Lect_7

LS4 Lect_7 - 1/25/10 Tet resistance Auxotroph revertant...

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1/25/10 1 Tet resistance Auxotroph revertant Bacteriophage resistance 5 bacteria 12 hours 10 7 bacteria 12 hours 12 hours 10 7 bacteria Media + tet E. Coli (10 7 tet s and 7 tet r ) Petri dish
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1/25/10 2 Complete media arg - E. coli 10 7 bacteria Minimal media Media + 10 10 T1 bacteriophage WT E. coli Ton s 10 7 bacteria Ton r : allele of Ton gene that makes bacteria resistant to T1 infection. Ton encodes the T1 receptor molecule. Complete Media with no penicillin WT E. coli 10 5 bacteria How can you isolate bacteria that have mutations that result in lethality? Need to isolate conditional lethals. 1. Penicillin selection. Penicillin causes defect in cell wall of bacteria, but only kills cells that are actively growing. WT E. coli + 100 auxotroph mutants X-rays Minimal media + penicillin plate
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3 Replica plate Selec%on vs screen. Selection: Establish conditions in which only the desired mutant will grow. Example: Select for Str r mutants on streptomycin. Penicillin selection. Genetic screen:
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LS4 Lect_7 - 1/25/10 Tet resistance Auxotroph revertant...

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