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Table of Contents
Why Do Genetics
Genetic Terms
More Terms
Basic Molelcular
Biology

More Basic Concepts
Screens
Selections
Mutation Frequency
Chemical Mutagenesis
Frameshift Mutation
DNA Repair
Mutation Summary
Detecting Mutants
Complex Mutation
Insertion Sequences
Compound Transposons
Complex Transposons
Models of
Transposition

Transposition Summary
Mutagenesis in vitro
Effects of Mutations
Complementation
Plasmids and
Conjugation

F Factor
Transformation
Transduction
Generalized
Transduction

Specialized
Transduction

Complementation
Mapping
Two Factor Crosses
Deletion Mapping
Other Mapping Methods
Strain Construction
Inverse Genetics
Gene Isolation
Characterization of
Clones

Sequence Data
General Approaches
Fusions
Supression
Final Summary
Problem Set 1
Problem Set 2


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A general approach to the analysis of a bacterial system

©2000 written by Gary Roberts, edited by Timothy Paustian, University of Wisconins-Madison

IX D. A GENERAL APPROACH TO THE GENETIC ANALYSIS OF A BACTERIAL SYSTEM

Thus far, the text has considered both good and bad approaches to solving genetic problems. The section following is a suggestion as to what elements one might want to use in the best of all possible worlds for the genetic analysis of a new bacterial system. The tools needed might be:

  1. A system of mutagenesis in vivo, presumably employing both chemical and transposon mutagenesis.

  2. A good screening system for the phenotype(s) of interest. For example, the identification of appropriately suboptimally supplemented plates would be useful. The requirement for "plates" or solid media is important since it allows examination of hundreds of isolated, potentially different mutants on each plate. You cannot examine enough different isolates in separate liquid cultures.

  3. An enrichment scheme for the phenotypes of interest.

  4. A system for genetic transfer in and out of the strain in vivo: this would typically entail conjugative plasmids capable of moving into the strain as well as plasmids capable of replicating within the strain.

  5. Methodology for transformation allowing the use of all manner of replicons constructed, manipulated or mutagenized in vitro.

  6. Niceties for studying regulation include the ability to assay reporter genes fused to a gene of interest. This can be powerful in assaying regulation and in demanding altered regulatory mutants (see Section XI).

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