An improved selection method for λlacZ- phages based on galactose sensitivity

Edwin J. Mientjes, Joost H M van Delft, Brenda M. op't Hof, Jan A. Gossen, Jan Vijg, Paul H M Lohman, Robert A. Baan

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

The determination of the lacZ mutant frequency in λgt10lacZ phage vectors isolated from the transgenic mouse strain 40.6 (MutaMouse), requires the screening of large numbers of phages on β-galactosidase activity. Existing methods rely on distinguishing a few white plaques on X-gal containing plates amongst a multide of blue ones which is both time-consuming and expensive. The new screening method described here employs the galactose sensitive Escherichia coli C lacZ recA galE strain into which a multicopy plasmid has been introduced, which results in over-expression of the galK and galT genes. In the presence of phenyl-β-d-galactopyranoside, a substrate for β-galactosidase, this leads to the suppression of λlacZ+ phage propagation without affecting the ability of λlacZ- phages to form plaques. With this method it is possible to screen 1.5×106 phages on a single 9-cm Petri dish. Furthermore, the need for blue/white screening has been eliminated.

Original languageEnglish (US)
Pages (from-to)67-69
Number of pages3
JournalTransgenic Research
Volume3
Issue number1
DOIs
StatePublished - Jan 1994
Externally publishedYes

Fingerprint

selection methods
Galactose
galactose
bacteriophages
Bacteriophages
galactosidases
Galactosidases
screening
phage vectors
plasmids
methodology
Transgenic Mice
genetically modified organisms
Escherichia coli
Plasmids
mutants
mice
genes
Genes

Keywords

  • λgt10lacZ phage
  • 40.6 transgenic mouse
  • galE
  • MutaMouse
  • mutant selection

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Genetics
  • Biotechnology
  • Plant Science
  • Food Science

Cite this

Mientjes, E. J., van Delft, J. H. M., op't Hof, B. M., Gossen, J. A., Vijg, J., Lohman, P. H. M., & Baan, R. A. (1994). An improved selection method for λlacZ- phages based on galactose sensitivity. Transgenic Research, 3(1), 67-69. https://doi.org/10.1007/BF01976029

An improved selection method for λlacZ- phages based on galactose sensitivity. / Mientjes, Edwin J.; van Delft, Joost H M; op't Hof, Brenda M.; Gossen, Jan A.; Vijg, Jan; Lohman, Paul H M; Baan, Robert A.

In: Transgenic Research, Vol. 3, No. 1, 01.1994, p. 67-69.

Research output: Contribution to journalArticle

Mientjes, EJ, van Delft, JHM, op't Hof, BM, Gossen, JA, Vijg, J, Lohman, PHM & Baan, RA 1994, 'An improved selection method for λlacZ- phages based on galactose sensitivity', Transgenic Research, vol. 3, no. 1, pp. 67-69. https://doi.org/10.1007/BF01976029
Mientjes EJ, van Delft JHM, op't Hof BM, Gossen JA, Vijg J, Lohman PHM et al. An improved selection method for λlacZ- phages based on galactose sensitivity. Transgenic Research. 1994 Jan;3(1):67-69. https://doi.org/10.1007/BF01976029
Mientjes, Edwin J. ; van Delft, Joost H M ; op't Hof, Brenda M. ; Gossen, Jan A. ; Vijg, Jan ; Lohman, Paul H M ; Baan, Robert A. / An improved selection method for λlacZ- phages based on galactose sensitivity. In: Transgenic Research. 1994 ; Vol. 3, No. 1. pp. 67-69.
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