[Plant-education] a-amylase expression

Carl Pike via plant-ed%40net.bio.net (by carl.pike from fandm.edu)
Mon Apr 21 22:15:19 EST 2008

I'm interested in the answer to Bob's question too, but would also 
appreciate any suggestions on a project of this sort that could be 
expanded into a multi-week investigation, including Westerns perhaps, 
in which students could hypothesize about the impact of some treatment 
on expression of the protein.  Maybe also link to measuring activity of 
enzyme in parallel.

Carl Pike
Department of Biology
Franklin and Marshall College

On Apr 21, 2008, at 5:57 PM, Bob Wise wrote:

> Dear Plant Edders,
> I need help.  One of the exercises I use in my plant physiology 
> teaching lab is the standard old expression of amylases during wheat 
> seed germination.  We take seeds germinated for 0 to 3 days, extract 
> total soluble protein and then run SDS-PAGE.
> The goal is to monitor the increase in the abundance of a-amylase 
> during germination.  The problem is that the bands don't get any 
> darker with germination time.  I have repeated the lab several times, 
> and changed to fresh seeds twice, yet I still cannot get an increase 
> in a-amylase expression.  The seeds are perfectly viable and work 
> quite well in a starch-digestion assay that forms the basis of a 
> separate lab exercise.
> Has anyone had success with this one?  Carol Reiss, are you out there?
> On the other hand, if this one is a bust, does anyone have a lab that 
> uses SDS-PAGE to show differential expression of plant proteins?  How 
> about a light vs. dark treatment to look for the increase in LHC 
> proteins during greening?  All I need is an electrophoresis lab that 
> clearly demonstrates protein synthesis during some plant developmental 
> or physiological process.  I would rather not do Westerns, due to time 
> constraints.
> Bob
> -- 
> Robert R. Wise
> Dept. of Biology
> UW Oshkosh
> 800 Algoma Blvd
> Oshkosh, WI 54901
> (920) 424-3404 (tel)
> (920) 424-1101 (fax)
> Have a look at The Structure and Function of Plastids at 
> http://www.life.uiuc.edu/govindjee/newbook/Vol%2023.html
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