Methods for monitoring multiple gene expression
View Patent ↗The present invention relates to methods for monitoring differential expression of a plurality of genes in a first filamentous fungal cell relative to expression of the same genes in one or more second filamentous fungal cells using microarrays containing Trichoderma reesei ESTs or SSH clones, or a combination thereof. The present invention also relates to computer readable media and substrates containing such array features for monitoring expression of a plurality of genes in filamentous fungal cells.
1. A method for monitoring differential expression of a plurality of genes in a first filamentous fungal cell relative to expression of the same genes in one or more second filamentous fungal cells, comprising:
(a) adding a mixture of detection reporter-labeled nucleic acids isolated from the first and one or more second filamentous fungal cells to a substrate containing an array of Trichoderma reesei ESTs or SSH clones, or a combination thereof, selected from the group consisting of SEQ ID NOs. 1-1188, under conditions where the nucleic acids hybridize to complementary sequences of the ESTs or SSH clones, or a combination thereof, in the array, wherein the nucleic acids from the first filamentous fungal cell and the one or more second filamentous fungal cells are labeled with a first detection reporter and one or more different second detection reporters, respectively; and
(b) examining the array under conditions wherein the relative expression of the genes in the first and one or more second filamentous fungal cells is determined by the observed detection signal of each spot in the array in which (i) the Trichoderma reesei ESTs or SSH clones, or a combination thereof, in the array that hybridize to the nucleic acids obtained from either the first or the one or more second filamentous fungal cells produce a distinct first detection signal or one or more second detection signals, respectively, and (ii) the Trichoderma reesei ESTs or SSH clones, or a combination thereof, in the array that hybridize to the nucleic acids obtained from both the first and one or more second filamentous fungal cells produce a distinct combined detection signal;
wherein the hybridization conditions are selected from the group consisting of very low, low, low-medium, medium, medium-high, high, and very high stringency conditions.
2. The method of claim 1 , wherein one or more of the first and one or more second filamentous fungal cells are selected from the group consisting of an Acremonium, Aspergillus, Aureobasidium, Bjerkandera, Ceriporiopsis, Coprinus, Coriolus, Cryptococcus, Filibasidium, Fusarium, Humicola, Magnaporthe, Mucor, Myceliophthora, Neocallimastix, Neurospora, Paediomyces, Penicillium, Phanerochaete, Phlebia, Piromyces, Pleurotus, Schizophyllum, Talaromyces, Thermoascus, Thielavia, Tolypodadium, Trametes, and Trichoderma cell.
3. The method of claim 1 , wherein the first and one or more second filamentous fungal cells are the same cell.
4. The method of claim 1 , wherein the first and one or more second filamentous fungal cells are Trichoderma reesei cells.
5. The method of claim 1 , wherein the first and one or more second filamentous fungal cells are different cells.
6. The method of claim 1 , wherein the detection reporter is fluorescence.
7. The method of claim 1 , wherein the differential expression of a plurality of genes in a first filamentous fungal cell relative to expression of the same genes in one or more second filamentous fungal cells is a result of growth of the first filamentous fungal cell on glucose and growth of the one or more second filamentous fungal cells on a cellulose, hemicellulose, or corn stover source.
8. The method of claim 1 , wherein the differential expression of a plurality of genes in a first filamentous fungal cell relative to expression of the same genes in one or more second filamentous fungal cells is a result of one or more different phenotypes.