have lab, need enzyme

Jon Monroe monroejd at jmu.edu
Thu Oct 9 10:35:38 EST 1997


David Starrett wrote: (some details omitted)

>Actually there is one that might work well.  Beta-galactosidase... There is
>easy PNP assay, there are generally 3-4 isozymes expressed in many tissues...

In what ways are they distinguishable besides pI?  Can you give us a recent
reference?  I see your abstract for the ASPP meeting
(http://www.sheridan.com/aspp/abs/46/1073.html)...  Good luck getting the
full length clone!

>it is compartmentalized within in the cell, and even cell wall.

Where is the form that is inside the membrane?  Vacuolar forms of
hydrolases seem to be very similar to apoplastic forms.

>In tomatoes, all isozymes appear to be
>glycosylated, but at different levels as Con-A can be used to fractionate
>the isozymes, to a degree anyways.

I was assuming that the Con-A column was an all or none process.  Can you
explain how you use it to fractionate glycosylated forms?

>  I am familiar with this stuff as it is my research area (and has been for
>5 years).  I adapted the extraction of both enzymes to my PP class here.
>The students have gotten so into it that half way through the current
>semester we have decided to drop some of the demo type experiments and
>carry these enzymes through a number of labs.

Great!  I'm hoping to head toward the same outcome...

>  Glad to give any more details, suggestions, etc.
>  Good luck,
>
>    Dave Starrett

Thanks!

Jon

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