[Plant-education] a-amylase expression

Bob Wise via plant-ed%40net.bio.net (by wise from uwosh.edu)
Mon Apr 21 16:57:03 EST 2008


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.

TIA

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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