Research MethodsResearch Methods kdorfman Thu, 10/20/2011 - 14:41
Pat's research methods course
Projects for Research MethodologyProjects for Research Methodology margaret Thu, 10/27/2011 - 20:50
Goals: determine the contribution of one (or more) spindle--associated protein(s) to mitosis in drosophila S2 cells (controls) and S2 cells in which the centrosome pathway is inhibited.
A. Culture S2 cells (untreated controls) and cells depleted of SAS--4 to block the centrosome pathway for spindle formation. Examine spindle formation for both conditions.
B. Select one or more Spindle associated proteins (list below), and deplete the protein from control cells and cells lacking SAS--4. Examine spindle formation for both conditions.
C. As an alternative to depletion of SAS--4, examine spindle formation in cells treated to disassemble microtubules and then allowed to reassemble microtubules (an assay to examine microtubule formation at chromosomes and centrosomes separately).
D. generate model(s) to explain your observations and test using additional depletions or manipulations.
Rationale: Many proteins have been shown to have a role in spindle formation, but how they contribute to spindle formation when the centrosome pathway is inhibited has not been examined.
Reading: Goshima et al Science 2007. Genes needed for mitosis in drosophila cells; also download the supplemental material for this paper. Goshima et al, JCB Mitotic motors paper Proteins.