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

254 Citations (Scopus)

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

Fingerprint

Genetic Engineering
Drug Resistance
Operon
Genetic Complementation Test
Chromosome Duplication
F Factor
Insertional Mutagenesis
Gene Fusion
Mutagenesis
Bacteriophages
Genetic Recombination
Chromosomes
Maintenance
Genome
Bacteria
Phenotype
Genes

ASJC Scopus subject areas

  • Virology

Cite this

Genetic engineering in Vivo using translocatable drug-resistance elements. New methods in bacterial genetics. / Kleckner, Nancy; Roth, John; Botstein, David.

In: Journal of Molecular Biology, Vol. 116, No. 1, 15.10.1977, p. 125-159.

Research output: Contribution to journalArticle

Kleckner, Nancy ; Roth, John ; Botstein, David. / Genetic engineering in Vivo using translocatable drug-resistance elements. New methods in bacterial genetics. In: Journal of Molecular Biology. 1977 ; Vol. 116, No. 1. pp. 125-159.
@article{d8743cf0a38d46e59dc1326c176f0d24,
title = "Genetic engineering in Vivo using translocatable drug-resistance elements. New methods in bacterial genetics",
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.",
author = "Nancy Kleckner and John Roth and David Botstein",
year = "1977",
month = "10",
day = "15",
doi = "10.1016/0022-2836(77)90123-1",
language = "English (US)",
volume = "116",
pages = "125--159",
journal = "Journal of Molecular Biology",
issn = "0022-2836",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Genetic engineering in Vivo using translocatable drug-resistance elements. New methods in bacterial genetics

AU - Kleckner, Nancy

AU - Roth, John

AU - Botstein, David

PY - 1977/10/15

Y1 - 1977/10/15

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0017742999&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0017742999&partnerID=8YFLogxK

U2 - 10.1016/0022-2836(77)90123-1

DO - 10.1016/0022-2836(77)90123-1

M3 - Article

C2 - 338917

AN - SCOPUS:0017742999

VL - 116

SP - 125

EP - 159

JO - Journal of Molecular Biology

JF - Journal of Molecular Biology

SN - 0022-2836

IS - 1

ER -