L2_3_31_11

L2_3_31_11 - READING FOR TUESDAY Ch. 3 125-142; 153-159;...

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Unformatted text preview: READING FOR TUESDAY Ch. 3 125-142; 153-159; Ch. 10 ALL TA ofce hours are posted on the web First problem set will be available on the web by tomorrow Podcasting is happening so Tuesdays lecture is on the web at media services. Please tell me if there are any problems Techniques in Cell Biology Making extracts Protein fractionation by size, Cell culture to test fractions Visual cells by phase contrast PCR to identify archaea FACS to purify archaea More chromatography, example of ion exchange, to purify active protein Structural analysis by X-ray crystallography to get info Chemical biology to synthesize similar small molecules that could act the same way. . Suppose you are a marine biologist that searches for new natural products that could be useful anti-cancer drugs. You are diving at a coral reef off of Tahiti, and you notice an unusual sponge that you have never seen before. You collect a sample, take it back to your lab, make an extract, and find that it contains a compound that kills cancer cells, but not normal cells. You are very excited and want to figure out where this compound comes from. Because you are a good marine biologist, you suspect that the compound actually comes from a micro-organism that has a symbiotic relationship with the sponge. You examine the sponge under a microscope and confirm that there are micro- organisms that are part of the sponge. You want to purify this cancer fighting compound. How do you do this? Phil Crews and his team http://chemistry.ucsc.edu/mnpr/expeditions.htm Fiji Hawai'i Madagascar Papua New Guinea US Virgin Islands Tonga Vanuatu Venezuela 1) Had a marvelous working vacation in Tahiti. 2) You collect a sample, take it back to your lab, make an extract and separate proteins based on size. Tools in cell biological research- Detergents and membrane solubilization Detergents Membrane solubilization with detergents Triton X-100 (a non-ionic detergent) dissolves membranes and solubilizes membrane proteins without affecting their structure/ function. SDS (an ionic detergent) dissolves membranes and denatures protein structure. You can also break open cells by osmotic shock, ultrasonic vibration or a blender + + The centrifuge is a key biochemical tool If you want to fractionate a cell extract, you centrifuge at different velocities. If you want to isolate a cell, its nuclei or cytoskeleton, you do a low speed spin Cell Fractionation Low speed pellet (1,000 x g) 10 minutes If you want to fractionate a cell to isolate an organelle, you do a medium speed spin Cell Fractionation (20,000 x g) 20 minutes Medium speed pellet If you want to fractionate a cell to isolate smaller organelles, you do a high speed spin....
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This note was uploaded on 02/06/2012 for the course BIO 110 taught by Professor Rexach during the Fall '08 term at UCSC.

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L2_3_31_11 - READING FOR TUESDAY Ch. 3 125-142; 153-159;...

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