Determination of mRNA half-lives in Candida albicans using thiolutin as a transcription inhibitor

Bessie W. Kebaara, Lindsey E. Nielsen, Kenneth W. Nickerson, Audrey L. Atkin

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

Abstract

A method for determining mRNA half-lives in the polymorphic fungus Candida albicans is described. It employs growth in a defined medium, the inhibition of transcription with thiolutin (10-20 μg/mL), and quantitative Northern blotting. The method is effective for the A72, SC5314, and CAI-4 strains of C. albicans, and for mRNAs that have a wide variety of decay rates and steady-state abundances. The range of half-lives detected (from 4-168 min) shows that this method is effective for mRNAs with widely varying half-lives. The mRNA decay rates obtained are compared with those for orthologous mRNAs from Saccharomyces cerevisiae. This procedure should work for a broad range of C. albicans strains and can be adapted to other fungal species.

Original languageEnglish (US)
Pages (from-to)894-899
Number of pages6
JournalGenome
Volume49
Issue number8
DOIs
StatePublished - Aug 2006

Keywords

  • 18S rRNA
  • ACT1
  • ADH1
  • Comparative mRNA stability
  • EFG1
  • PGK1
  • mRNA decay

ASJC Scopus subject areas

  • Biotechnology
  • Molecular Biology
  • Genetics

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