Kebaara BW, Nielsen LE, Nickerson KW, Atkin AL.Determination
of mRNA half-lives in Candida albicans using thiolutin as a
transcription inhibitor.
Genome. 2006 Aug;49(8):894-9.
School of Biological Sciences, University of
Nebraska-Lincoln, NE 68588-0666, USA.
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.
PMID: 17036064
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