Mini-Mu technology in Salmonella typhi: Isolation of stable MudJ operon fusions by cis complementation

I. Contreras, V. Obreque, B. Tesser, G. C. Mora

Resultado de la investigación: Article

2 Citas (Scopus)

Resumen

This paper describes a method to achieve stable MudJ insertions in the Salmonella typhi Ty2 chromosome. The method is a modification of the genetic complementation system described previously for Salmonella typhimurium, which consists in placing the defective transposon (MudJ) near the transposase genes of a helper Mu phage on a single DNA fragment. This fragment is then introduced into a new bacterial host by means of P22 transduction. We constructed a S. typhi strain which carries MudJ and the Mu helper phage in the chromosome. This strain was induced to lytic growth and the lysate was used to infect S. typhi Ty2. The frequency of mutation was 2.0 x 10-6 mutants per recipient bacterium. Superinfection with the Mu helper phage was about 1%. To determine the number of MudJ insertions, several mutants were subjected to Southern blot analysis. From a total of 25 mutants analyzed, only 4 contained more than one insertion. Our procedure compares well with the method described previously for the S. typhimurium-P22 system and can be applied to other Mu sensitive bacteria.

Idioma originalEnglish
Páginas (desde-hasta)233-239
Número de páginas7
PublicaciónBiological Research
Volumen27
N.º3-4
EstadoPublished - 1994

Huella dactilar

Salmonella Typhi
Salmonella typhi
Bacteriophages
Salmonella
Bacteriophage mu
operon
Operon
bacteriophages
Fusion reactions
Chromosomes
Technology
Salmonella Typhimurium
mutants
Bacteria
Salmonella typhimurium
genetic complementation
Transposases
chromosomes
bacteria
transposons

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Citar esto

Contreras, I., Obreque, V., Tesser, B., & Mora, G. C. (1994). Mini-Mu technology in Salmonella typhi: Isolation of stable MudJ operon fusions by cis complementation. Biological Research, 27(3-4), 233-239.
Contreras, I. ; Obreque, V. ; Tesser, B. ; Mora, G. C. / Mini-Mu technology in Salmonella typhi : Isolation of stable MudJ operon fusions by cis complementation. En: Biological Research. 1994 ; Vol. 27, N.º 3-4. pp. 233-239.
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abstract = "This paper describes a method to achieve stable MudJ insertions in the Salmonella typhi Ty2 chromosome. The method is a modification of the genetic complementation system described previously for Salmonella typhimurium, which consists in placing the defective transposon (MudJ) near the transposase genes of a helper Mu phage on a single DNA fragment. This fragment is then introduced into a new bacterial host by means of P22 transduction. We constructed a S. typhi strain which carries MudJ and the Mu helper phage in the chromosome. This strain was induced to lytic growth and the lysate was used to infect S. typhi Ty2. The frequency of mutation was 2.0 x 10-6 mutants per recipient bacterium. Superinfection with the Mu helper phage was about 1{\%}. To determine the number of MudJ insertions, several mutants were subjected to Southern blot analysis. From a total of 25 mutants analyzed, only 4 contained more than one insertion. Our procedure compares well with the method described previously for the S. typhimurium-P22 system and can be applied to other Mu sensitive bacteria.",
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Contreras, I, Obreque, V, Tesser, B & Mora, GC 1994, 'Mini-Mu technology in Salmonella typhi: Isolation of stable MudJ operon fusions by cis complementation', Biological Research, vol. 27, n.º 3-4, pp. 233-239.

Mini-Mu technology in Salmonella typhi : Isolation of stable MudJ operon fusions by cis complementation. / Contreras, I.; Obreque, V.; Tesser, B.; Mora, G. C.

En: Biological Research, Vol. 27, N.º 3-4, 1994, p. 233-239.

Resultado de la investigación: Article

TY - JOUR

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AU - Tesser, B.

AU - Mora, G. C.

PY - 1994

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N2 - This paper describes a method to achieve stable MudJ insertions in the Salmonella typhi Ty2 chromosome. The method is a modification of the genetic complementation system described previously for Salmonella typhimurium, which consists in placing the defective transposon (MudJ) near the transposase genes of a helper Mu phage on a single DNA fragment. This fragment is then introduced into a new bacterial host by means of P22 transduction. We constructed a S. typhi strain which carries MudJ and the Mu helper phage in the chromosome. This strain was induced to lytic growth and the lysate was used to infect S. typhi Ty2. The frequency of mutation was 2.0 x 10-6 mutants per recipient bacterium. Superinfection with the Mu helper phage was about 1%. To determine the number of MudJ insertions, several mutants were subjected to Southern blot analysis. From a total of 25 mutants analyzed, only 4 contained more than one insertion. Our procedure compares well with the method described previously for the S. typhimurium-P22 system and can be applied to other Mu sensitive bacteria.

AB - This paper describes a method to achieve stable MudJ insertions in the Salmonella typhi Ty2 chromosome. The method is a modification of the genetic complementation system described previously for Salmonella typhimurium, which consists in placing the defective transposon (MudJ) near the transposase genes of a helper Mu phage on a single DNA fragment. This fragment is then introduced into a new bacterial host by means of P22 transduction. We constructed a S. typhi strain which carries MudJ and the Mu helper phage in the chromosome. This strain was induced to lytic growth and the lysate was used to infect S. typhi Ty2. The frequency of mutation was 2.0 x 10-6 mutants per recipient bacterium. Superinfection with the Mu helper phage was about 1%. To determine the number of MudJ insertions, several mutants were subjected to Southern blot analysis. From a total of 25 mutants analyzed, only 4 contained more than one insertion. Our procedure compares well with the method described previously for the S. typhimurium-P22 system and can be applied to other Mu sensitive bacteria.

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