[Chlamydomonas] lysis conditions

Josh Rappoport rappopj at mail.rockefeller.edu
Tue Jul 19 12:10:46 EST 2005


I am interested in making a Chlamy cell lysate for use in 
pulling-down binding partners to a GST-fusion protein expressed 
in bacteria and then bound to glutathione beads.
Thus, I would like to be able to get at the cytosolic proteins 
leaving behind the nuclei, chloroplasts, and the associated DNA.
Does anyone have a protocol for doing this?
I suppose it would start with getting rid of the cell wall, 
followed by disruption either mechanically and/or by mild 
detergents and finally ending with pelleting of insoluble 
material.
Thank you very much.

Josh

__________________________
Joshua Z. Rappoport PhD
Post-doctoral Fellow
The Rockefeller University
Laboratory of Cellular Biophysics
1230 York Avenue, Box 304
New York, NY 10021

Phone: 347-231-5289
Fax: 212-327-7543




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