Tour-de-Force

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Tour-de-Force: Interplay between Mitochondria and Cell Cycle Progression Fall 2007Although the majority of the studies on cyclin D effects on NRF-1 have been focused on cdk4, thisdoes not exclude cdk6 as another possible mediator. By using co-immunoprecipitation we can see whenwhich cyclin D isoform binds to which cdk.Possible role of other cdk’s in mediating regulation of NRF-1 and NRF-2 activityDue to the fact that it is cdk4 that is responsible for the phosphorylation of NRF-1 it would be interesting tosee if other cdk’s are also involved in this regulation during the cell cycle.We would therefore first like to check if these cdks can interact with NRF-1 and 2. In order to do this wewould like to do kinase assay using the AlphaScreen® PhosphoSensor Kits (PerkinElmer).SCI 332 Advanced Molecular Cell Biology Research Proposal 68
Tour-de-Force: Interplay between Mitochondria and Cell Cycle Progression Fall 2007Proposed CostsItem Year 1 Year 2 Year 3 Year 4 Total itemexpenditurePhD student 40,000 40,000 40,000 40,000 160,000Bench fee 10,000 10,000 10,000 10,000 40,000Antibodies 3,000 3,000Custom antibodies 1,500 1,500FACS probes 350 350Cell lines 600 600 600 600 2,400Phos-tools 650 650PCR primers 300 800 1,100PCR probes 150 150 300PCR First Strand Kit 150 150Taqman assay 1100 1,100Light Cycler 2.0 ® RT-PCR system10,000 10,000 15,000 35,000Nocodazole 250 250AlphaScreen®PhosphoSensor KitsMAPK p38 & ErkinhibitorssiRNA (custom siRNA& materials)260 260700 700800 800Unforeseen costs 2440Total per year 62,600 62,250 71,850 50,860Total expenditure 250,000TimelineActivity Year 1 Year 2 Year 3 Year 4PreparationSubtopic 1Subtopic 2Subtopic 3SCI 332 Advanced Molecular Cell Biology Research Proposal 69
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Tour-de-Force

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