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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