Characterization of color mutants in lacZ plasmid-based transgenic mice, as detected by positive selection

Martijn E T Dollé, Hans Jörg Martus, Maja Novak, Nathalie J. Van Orsouw, Jan Vijg

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

The plasmid-based transgenic mouse model, which uses the lacZ gene as the target for mutation, is sensitive to a wide range of in vivo mutations, ranging from point mutations to insertions and deletions extending far into the mouse genome. In this study, the nature of subtle lacZ mutations, which do not completely abolish β-galactosidase activity, as detected by positive selection, was investigated. These subtle mutants are called 'color mutants' due to their light blue staining on X-gal medium. Replating of color mutants and retransformation of plasmid DNA, purified from individual color mutants, resulted in the same phenotype as the original color mutant. The p-gal positive selection system tolerates ~ 10% of wild-type activity as indicated by spectrophotometric determination of β-galactosidase activity of individual color mutants. Restriction digestion and size separation of plasmid DNA revealed no visible change in the size of the plasmid in color mutants. Sequence analysis confirmed the presence of a point mutation in each lacZ gene of nine different color mutants. The results indicate that color mutants are caused neither by the presence of a mixture of wild-type and mutated lacZ plasmids within the same host cell nor by a mixture of cells within the original mutant colony which carry either wild-type or mutated lacZ plasmids. In addition, it was discovered that the mouse line studied harbors four polymorphic base changes among the integrated plasmid copies.

Original languageEnglish (US)
Pages (from-to)287-293
Number of pages7
JournalMutagenesis
Volume14
Issue number3
DOIs
StatePublished - 1999
Externally publishedYes

Fingerprint

Transgenic Mice
Plasmids
Color
Galactosidases
Lac Operon
Genes
Point Mutation
Mutation
Size separation
DNA
Ports and harbors
Sequence Analysis
Digestion
Genome
Staining and Labeling
Phenotype

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)
  • Toxicology
  • Health, Toxicology and Mutagenesis

Cite this

Characterization of color mutants in lacZ plasmid-based transgenic mice, as detected by positive selection. / Dollé, Martijn E T; Martus, Hans Jörg; Novak, Maja; Van Orsouw, Nathalie J.; Vijg, Jan.

In: Mutagenesis, Vol. 14, No. 3, 1999, p. 287-293.

Research output: Contribution to journalArticle

Dollé, Martijn E T ; Martus, Hans Jörg ; Novak, Maja ; Van Orsouw, Nathalie J. ; Vijg, Jan. / Characterization of color mutants in lacZ plasmid-based transgenic mice, as detected by positive selection. In: Mutagenesis. 1999 ; Vol. 14, No. 3. pp. 287-293.
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