Genetic engineering in Vivo using translocatable drug-resistance elements. New methods in bacterial genetics

Nancy Kleckner, John Roth, David Botstein

Research output: Contribution to journalArticle

256 Scopus citations

Abstract

A number of translocatable drug-resistance elements have recently been described which are able to insert themselves into a large number of different sites in prokaryotic genomes. These elements cause recognizable mutations when insertion occurs within a structural gene or an operon. Drug-resistance elements are also associated with other kinds of illegitimate recombination events, notably deletions and inversions. This paper summarizes uses to which these properties of translocatable drugr-esistance elements can be put in genetic manipulations of bacteria. Translocatable drug-resistance elements are useful in isolation of mutants (even where the mutant phenotype is not easily scored), in the construction of strains and other genetic manipulations (even when selection is difficult or impossible), in localized mutagenesis, in chromosomal mapping, in construction of Hfr strains with known origin and direction of chromosome transfer, in complementation tests, in construction of new F′ plasmids, in construction of new specialized transducing phages, in isolation of deletions with one or both endpoints specified, in construction of gene and operon fusions, and in the selection and maintenance of chromosomal duplications. Experiments are described which illustrate many of these techniques.

Original languageEnglish (US)
Pages (from-to)125-159
Number of pages35
JournalJournal of Molecular Biology
Volume116
Issue number1
DOIs
StatePublished - Oct 15 1977

ASJC Scopus subject areas

  • Virology

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