2. Academic Validation
  3. Determination of mRNA half-lives in Candida albicans using thiolutin as a transcription inhibitor

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

  • Genome. 2006 Aug;49(8):894-9. doi: 10.1139/g06-046.
Bessie W Kebaara 1,  Lindsey E Nielsen,  Kenneth W Nickerson,  Audrey L Atkin
Affiliations

Affiliation

  • 1 School of Biological Sciences, University of Nebraska-Lincoln, NE 68588-0666, USA.
PMID: 17036064 DOI: 10.1139/g06-046
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 microg/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.

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