Tour-de-Force

More documents

Recommendations

Info

Tour-de-Force: Interplay between Mitochondria and Cell Cycle Progression Fall 2007Proposed CostsSalary (PhD student):Bench-fee:40 000 EUR/year (total 160 000 EURO in 4 years)10 000 EUR/year (total 40 000 EURO in 4 years)Experiments andMaterials Year 1 Year 2 Year 3 Year 4General MaterialsCell lines €2.150 - - -Medium €400 €600 €600 €400Nocodazole €100 €100 €100 €100BrdU + antibodies €400 €400 €400 -Histone H3phosphorylation €500 €500 €500 €500Question 1Self-referencing system €23.000 - - -JC-1 mitochondrialdetection kit€200 - - -Question 2ROS probes €1.500 €1.500 - -Antioxidant enzyme kits - €800 - -Question 3Materials ROS level - - €400 -manipulationReal Time RT-PCR - - €500 -IP kinase assay - - - €1.000Antibodies - - - €5.00035 S pulse-chaseexperiment- - - €1.000Total: €5.250 €3.900 €2.500 €8.000€19.650For all machinery used during the proposed experiments, we will use the university as our resource forperforming these measurements. A total of € 242.650 funding is needed for the research proposal.SCI 332 Advanced Molecular Cell Biology Research Proposal 32
Tour-de-Force: Interplay between Mitochondria and Cell Cycle Progression Fall 2007Proposed TimelineFor the entire research, a four-year time span has been allotted. The estimated duration of thesubquestions in all cell lines and conditions is shown in this overview.Question 1: Are there cyclic fluctuations in mitochondrial metabolic activity that are coordinated with thecell cycle?The total time span of question 1, including data analysis, will be around six months. Measurement of oxygen consumption and BrdU-labelling:4 months Measurement of membrane potential and BrdU-labelling:2 monthsQuestion 2: Do ROS levels fluctuate throughout the cell cycle?Approximately 19 months will be allowed for question 2. Making and testing the Hyper and PF-1 probe: Measurement of cytosolic and mitochondrial superoxide and hydrogenperoxide levels: Measurement of peroxisomal hydrogen peroxide levels: Measurement of cytosolic superoxide and hydrogen peroxidewhile NADPH oxidase is inhibited: Control for laser-induced ROS production: Measurement of antioxidant activity: Data analysis:3 months5 months1 month2 months1 month3 months4 monthsQuestion 3: Is the G 2 /M transition controlled by redox-dependent regulation of cyclin-cdk complexes?The total time span of this question will probably cover eighteen months. Determination doses antioxidants, antimycin A and hydrogen peroxide: 4 months Cell cycle progression with FACS:2 months IP Kinase assay and Western blot:2 months Northern blotting and real time RT-PCR:3 months 35-S pulse-chase experiments:1 month Further co-immunoprecipitation:2 months Data analysis:4 monthsThis planning ensures that the last months of the four year time span are available for processing all thedata and combining it into a conclusive research. It also leaves some space for unforeseen events, suchas repetition of experiments.Question 1Question 2Question 3Total Data AnalysisYear 1 Year 2 Year 3 Year 4SCI 332 Advanced Molecular Cell Biology Research Proposal 33
Page 1 and 2: Tour-de-ForceInterplay between mito
Page 3 and 4: Tour-de-Force: Interplay between Mi
Page 31: Tour-de-Force: Interplay between Mi
Page 83 and 84:
Tour-de-Force: Interplay between Mi
Page 85 and 86:
Page 87 and 88:
Page 89 and 90:
Page 91 and 92:
Page 93 and 94:
Page 95 and 96:
Page 97 and 98:
Page 99 and 100:
Page 101 and 102:
Page 103 and 104:
Page 105 and 106:
Page 107 and 108:
Page 109 and 110:
Page 111 and 112:
Page 113 and 114:
Page 115 and 116:
Page 117 and 118:
show all

Tour-de-Force

You also want an ePaper? Increase the reach of your titles

Delete template?

Save as template?