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