Fechar
gene
Open Reading Frame (ORF)
mutation
allele
genotype
phenotype
If asking: What GENES cause this PHENOTYPE?
Selectable mutations
Non-selectable mutations
auxotroph
Selection
selection: pros and cons
Screen
Screen: pros and cons
Replica plating
essential gene
How to screen for a conditional loss of function mutant
mutation rate
spontaneous mutations
microlesions
macrolesions
Mutagenesis
Chemical Mutagens: Nucleotide Base Analogs
UV radiation
SOS response system
transposons
transposase
Engineered Transposons: Requirements
Why can't the inserted mini-Tn not move again?
advantage of Tn mutagenesis
genetic exchange
horizontal gene transfer
transformation (natural)
Griffith experiment
F plasmid
homologous recombination
competence
Plasmid
conjugation
tra regions of F plasmid
oriT region
oriV region
transposons/IRs in F plasmid
Why would we use bacterial conjugation?
transduction
transducing particle
transductants
Why use transduction?
How to combine two mutations in the chromosome of one bacterial strain
Why would we want to express a particular gene from a plasmid in a bacterial host?
(think: how could we understand a particular gene's function?)
complementation
How to find affected gene when using UV or chemical mutagenesis
When to use RANDOM mutagenesis
When to use TARGETED mutagenesis
targeted gene disruption
How to transform some competent bacterium with a knockout plasmid
When does targeted gene disruption NOT work?
How would we learn the null phenotype of an essential gene?
inducible promoters
Forward genetics
Reverse genetics
To mutate a known gene using a transposon
Transposon Mutagenesis vs Targeted gene disruption
