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 filamentous fungal expressed sequenced tags. The present invention also relates to filamentous fungal expressed sequenced tags and to computer readable media and substrates containing such expressed sequenced tags 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 fluorescence-labeled nucleic acids isolated from the filamentous fungal cells to a substrate containing an array of Fusarium venenatum ESTs comprising SEQ ID NOs. 1–3770, under conditions where the nucleic acids hybridize to complementary sequences of the ESTs in the array, wherein the nucleic acids from the first filamentous fungal cell are labeled with a first fluorescent reporter and the one or more second filamentous fungal cells are labeled with one or more different second fluorescent reporters; and
(b) examining the array by fluorescence under fluorescence excitation conditions wherein the relative expression of the genes in the filamentous fungal cells is determined by the observed fluorescence emission color of each spot in the array in which (i) the fluorescence-labeled nucleic acids obtained from the first filamentous fungal cell that are hybridized to the ESTs in the array produce a distinct first fluorescence emission color and the fluorescence-labeled nucleic acids obtained from the one or more second filamentous fungal cells that are hybridized to the ESTs in the array produce a distinct second fluorescence emission color, and (ii) the fluorescence-labeled nucleic acids obtained from both the first and the one or more second filamentous fungal cells that are hybridized to the ESTs in the array produce a distinct combined fluorescence emission color.
2. The method of claim 1 , wherein one or more of the filamentous fungal cells are selected from the group consisting of an Acremonium, Aspergillus, Fusarium, Humicola, Mucor, Myceliophthora, Neurospora, Penicillium, Thielavia, Tolypocladium, and Trichoderma cell.
3. The method of claim 1 , wherein the two or more filamentous fungal cells are the same cell.
4. The method of claim 1 , wherein the two or more filamentous fungal cells are Fusarium venenatum cells.
5. The method of claim 1 , wherein the two or more filamentous fungal cells are Aspergillus niger cells.
6. The method of claim 1 , wherein the two or more filamentous fungal cells are Aspergillus oryzae cells.
7. The method of claim 1 , wherein the two or more filamentous fungal cells are different cells.
8. The method of claim 1 , 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.
9. The method of claim 1 , wherein the array comprises the EST of SEQ ID NO: 6 and one or more ESTs selected from the group consisting of SEQ ID Nos: 1–5 and 7–3770.