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Experiment: Targets of the mci genes.
Experiment Description
NASCArrays Experiment Reference Number: NASCARRAYS-74
In Antirrhinum, the equivalent mutant to the Arabidopsis cuc1 cuc2 double is called cup. We have cloned CUP and shown that it encodes a NAC-domain transcription factor homologous to CUC1 and CUC2. Yeast two-hybrid analysis shows that CUP interacts with TIC, an Antirrhinum TCP transcription factor. Moving back to Arabidopsis, the closest homologues to TIC encode TCP factors TCP13 and TCP14 which, we have now shown, also interact in two-hybrid experiments with CUC1 and CUC2. We have identified insertions in both TCP13 and TCP14. CUP, CUC1 and CUC2 play a role in the establishment of boundaries between lateral organs. As evolutionarily conserved interactors, we expect TCP13 and TCP14 to act in the same process. Homozygous tcp13 mutant flowers show mixed cell identity (mci) with the boundaries of organ identity out of register with those of physical organ development. tcp 14 mutants show a very weak phenotype, but the double heterozygote is identical to the tcp13 homozygote. The double homozygote is underway at the moment and this application takes into account the time required to generate these plants.The aim of this microarray experiment is to identify targets of TCP13 and TCP14. We propose to compare expression levels in WT, tcp13, tcp14, tcp13/+ tcp14/+, and tcp13 tcp14 plants. Although we initially propose only duplicate experiments for each mutant, requiring a total of 10 chips, the experimental material will serve as internal replicates, since we expect to see different degrees of inactivation/activation of at least a core of conserved genes. The fact that all of the mutant combinations observed so far produce flowers with the appropriate cell types present, but in a different position, suggests that, unlike in homeotic mutants, very similar sets of floral genes will be present in each sample. This gives us hope that we might find a smaller number of genes with altered expression. Our tissue samples will be wild type and mutant inflorescences and, as such, will contain a variety of flowers at different stages of development. By taking a large number of flowers and buds we hope to overcome any sampling errors. Although we have requested 10 chips - for duplicate analyses of WT, both single mutants, the heterozygote and the double mutant - it remains a formal possibility that the double homozygote is inviable, or produces no inflorescence. In this unlikely eventuality we would only submit 8 of the 10 samples.
About the ExperimenterName: | Dr Brendan Davies |
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Head of Lab Name: | Dr Brendan Davies |
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Lab:
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Institute:
| University of Leeds |
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Address: | Centre for Plant Sciences School of Biology University of Leeds Leeds
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Postcode:
| LS2 9JT |
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Country:
| UK |
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| Telephone Number:
| 0113 343 3123 |
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Fax Number:
| 0113 343 3144 |
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are free to do whatever you please with it. If you intend to publish
work based on any of this data, please acknowledge us, contact the
experimenter above, and either acknowledge them or use them as
co-authors in the work.
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| About this ExperimentExperiment Type:
| genetic_modification_design |
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Number of Slides: | 8 |
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| Experimental Parameters:
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parameter | gene_knock_out |
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Quality Control Measures Taken:
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no-plants-pooled | 10 |
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References:
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| Other Information:
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ArrayExpress Accession | E-NASC-19 |
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Slides in this Experiment
Hybridisation Set: Davies_genome
Slide: Davies_1-1_Con_Rep1_ATH1 | | |
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Tissue:
| inflorescence |
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Diseased:
| Normal |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Davies_1-2_Con_Rep2_ATH1 | | |
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Tissue:
| inflorescence |
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Diseased:
| Normal |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Davies_1-3_T13_Rep1_ATH1 | | |
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Tissue:
| inflorescence |
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Diseased:
| Normal |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Davies_1-4_T13_Rep2_ATH1 | | |
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Tissue:
| inflorescence |
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Diseased:
| Normal |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Davies_1-5_T14_Rep1_ATH1 | | |
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Tissue:
| inflorescence |
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Diseased:
| Normal |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Davies_1-6_T14_Rep2_ATH1 | | |
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Tissue:
| inflorescence |
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Diseased:
| Normal |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Davies_1-7_Het_Rep1_ATH1 | | |
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Tissue:
| inflorescence |
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Diseased:
| Normal |
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Other Information:
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Protocols for BioSource 1 |
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Slide: Davies_1-8_Het_Rep2_ATH1 | | |
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Tissue:
| inflorescence |
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Diseased:
| Normal |
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Other Information:
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Protocols for BioSource 1 |
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