yeast transformation - how to make a thermotolerant yeast (Mar/02/2009 )

hi guys!

im currently doing a research proposal for my biotechnology course and im interested in making a thermotolerant yeast but then i dont know where to start

i hope you can help me....


thanks a lot

janice

Janice on Mar 2 2009, 03:43 AM said:

Directed evolution?
Mutagen treatment, grow at 39 degree C, repeat the cycles. You may want to elevate the temperature.

WHR on Mar 2 2009, 04:57 AM said:

hi whr!

thanks for your reply. what mutagens can i use? can i just use plasmids for transformation?


thanks

Short wave UV exposure is probably easiest and safest (don't get a sunburn). You will likely have to try a range of exposure times to find the sub-lethal exposures which cause mutations but not too much death. If you have a Stratagene membrane cross linker (Stratalinker) that would be ideal, but any source of 254 nm UV would work.

As phage suggested. Besides, there are many chemical mutagens.
I think thermo tolerance is affected by multiple genes. Overexpression of certain heat shock proteins may have little effects. (?)