In a collaboration with the Advanced Center for Genome Technology (ACGT, University of Oklahoma), we have sequenced 13,000 cDNA clones from two time-of-day specific libraries (morning and evening library) generating approximately 20,000 sequences. Contig analysis allowed the identification of 445 unique ESTs and 986 ESTs present in multiple cDNA clones. For about 50 % of the sequences (710 of 1,431), significant matches to sequences in the NCBI database (of known or unknown function) were detected. You can view the contigs or single clones and their putative homologues here. You can also search the EST sequence data using BLAST. We hybridized Northern blots with probes derived from 26 clones chosen from contigs identified by multiple cDNA clones and EST sequences. Using these sequences, the representation of genes among the morning and evening sequences, respectively, in most cases does not reflect their expression patterns over the course of the day. Nevertheless, we were able to identify four new clock-controlled genes. Based upon these data we predict that a significant proportion of the expressed Neurospora genes may be regulated by the circadian clock. The mRNA levels of all four genes peak in the subjective morning as is the case with previously identified ccgs.
The data are published in:
Zhu H, Nowrousian M, Kupfer D, Colot HV, Berrocal-Tito G, Lai H, Bell-Pedersen D, Roe BA,
Loros JJ, Dunlap JC (2001). Analysis of ESTs from two starvation, time of day-specific
libraries of Neurospora crassa reveals novel clock-controlled genes.
Genetics 157: 1057-1065