Determination of mRNA half-lives in Candida albicans using thiolutin as a transcription inhibitor.
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.
Kebaara
Bessie W
BW
School of Biological Sciences, University of Nebraska-Lincoln, NE 68588-0666, USA.
Nielsen
Lindsey E
LE
Nickerson
Kenneth W
KW
Atkin
Audrey L
AL
D001483:Base Sequence; D002176:Candida albicans / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D017931:DNA Primers / Q000235:genetics; D006207:Half-Life; D011760:Pyrrolidinones / Q000494:pharmacology; D012331:RNA, Fungal / Q000235:genetics* / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects
D017931:DNA Primers; D011760:Pyrrolidinones; D012331:RNA, Fungal; D012333:RNA, Messenger; C006361:acetopyrrothine