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yeast transformation


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#1 Janice

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Posted 02 March 2009 - 03:43 AM

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

#2 WHR

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Posted 02 March 2009 - 04:57 AM

View PostJanice, on Mar 2 2009, 03:43 AM, said:

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

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

#3 Janice

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Posted 02 March 2009 - 06:40 PM

View PostWHR, on Mar 2 2009, 04:57 AM, said:

View PostJanice, on Mar 2 2009, 03:43 AM, said:

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

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


hi whr!

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


thanks

#4 phage434

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Posted 02 March 2009 - 08:31 PM

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.

#5 WHR

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Posted 02 March 2009 - 11:03 PM

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. (?)




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