Notes

Slide Show

Outline

1	EB1 proteins in Arabidopsis A colorful overview
2	The Cytoskeleton
3	Cytoskeletal Functions Include: Determination of cell shape Cell Division Cell Elongation Organelle movement Cell signaling
4	EB1 family proteins Evolutionarily conserved Functions + end microtubule (MT) binding Centrosome binding in animals cells MT dynamics and organization EB1 in Arabidopsis 3 homologs: EB1a (831 base pairs) EB1b (882 bp) EB1c (990 bp)
5	EB1 Protein Structure Structure Calponin homology (CH) domain MT binding domain C-terminal motif Long acidic tail, different from all other EB1 proteins Binding domain for additional protein? APC/p150glued /actin
6	Purpose Gain familiarity with basic lab techniques Develop a MT reporter to study the dynamics of the cytoskeleton Fusions with GFP, YFP, and CFP Truncated versions Binding domain Probe the significance of the CH domain Interaction with actin?
7	Techniques and Basic Procedures PCR-based cDNA cloning E. coli transformation with fusion constructs Transient expression in tobacco Leaf bombardment Fluorescent imaging with the confocal microscope
8	Full Length EB1a
9	Full Length EB1a cont.
10	Full Length EB1a cont.
11	Full Length EB1b
12	Full Length EB1b cont.
13	Full Length EB1b cont.
14	Full Length EB1b cont.
15	Full Length EB1c
16	Truncated EB1s
17	Comparing MT Reporters
18	Actin-MT Co-visualization
19	Co-visualization cont.
20	EB1b and Fimbrin
21	Future Endeavors Create stable Arabidopsis lines expressing the different fusions Study MT dynamics in stable, transformed cells Better co-visualization with actin
22	Acknowledgements and Multiple Thank-Yous The Noble Foundation Steve Rhines Sue Elison Christy Cheolmin Brittany Rex and Fuzzy IV