We have a grape breeding program here at our lab. I have been doing alot
of work with vitis (grape) and I find them very difficult to work with -
they have a lot of polyphenolics and acids in their leaf cells.
I am not suprised that there is some kind of interference with an enzymatic
assay as complex as Rubisco. What exactly are you trying to study? Can
you switch to measuring Chlorophyll, or some other type of assay?
When isolating DNA, I had to resort to doing a nuclear isolation first thus
protecting the DNA from all of the vacuole and cytoplasmic junk. I also
work with very young shoot tip leaves at the very beginning of the growing
I am sorry I don't have any ideas to help you with your specific assay, but
maybe my experiences will give you some ideas. I do wish you luck.
Molecular Markers Lab Technician
USDA Agricultural Research Service
2021 S. Peach Ave.
Fresno, CA 93727
e-mail: dbell at qnis.net
phone: 559 453-3170
fax: 559 453-3088
> From: marcelo <msegura at pregrado.ciencias.uchile.cl>
> To: methods at hgmp.mrc.ac.uk> Subject: Rubisco activty
> Date: Wednesday, January 24, 2001 1:05 PM
>> hello! I have a problem with measuring the activity of Ru in grape
> leaves. I am using an espectrophotometric assay with coupling enzymes of
> the Calvin cycle. As control we used spinach leaves and the methods
> Thank`s very much