A Tn10 derivative (T-POP) for isolation of insertions with conditional (tetracycline-dependent) phenotypes

Chad A. Rappleye, John R. Roth

Research output: Contribution to journalArticle

67 Citations (Scopus)

Abstract

A new Tn10-based transposon has been constructed and used to isolate insertion mutations with tetracycline-conditional phenotypes. Classes of mutants include conditional lethal mutations, conditional auxotrophs, and conditional mutants of the eut (ethanolamine utilization) operon. The described mutations were made with a new derivative of Tn10dTet that we have called Tn10d(T-POP). Others have noted that transposon Tn10dTet directs weak tetracycline-inducible transcripts out of both ends of the element into adjacent sequences. We have increased this level of outward transcription from Tn10dTet by selecting deletion mutations within the element that presumably remove transcription-termination signals. Insertion of the Tn10d(T-POP) element within an operon disrupts the target gene and makes expression of distal genes dependent on induction of outward transcription by tetracycline. Insertion mutations made with Tn10d(T-POP) can cause tetracycline-correctable conditional phenotypes based on expression of distal genes.

Original languageEnglish (US)
Pages (from-to)5827-5834
Number of pages8
JournalJournal of Bacteriology
Volume179
Issue number18
StatePublished - Sep 1997
Externally publishedYes

Fingerprint

Tetracycline
Phenotype
Insertional Mutagenesis
Operon
Gene Expression
Mutation
Ethanolamine
Sequence Deletion
Genes

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Immunology

Cite this

A Tn10 derivative (T-POP) for isolation of insertions with conditional (tetracycline-dependent) phenotypes. / Rappleye, Chad A.; Roth, John R.

In: Journal of Bacteriology, Vol. 179, No. 18, 09.1997, p. 5827-5834.

Research output: Contribution to journalArticle

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