Date: Tue, 20 Jan 1998 04:59:16 -0700 From: cfrazie@unm.edu (Chris Frazier) To: cp@opus.hpl.hp.com Message-Id: <aabcdefg249$foo@default> Subject: Re: Nep project/ Electrophoresis
CPfolk,
First off, I apologize for inadvertantly sending a personal
message to Tan Wee Kiat to the list by mistake. I hate it when that
happens. In any case, as my email mentioned, I will be out in Singapore
and Peninsular Malaysia from about March 1 to the end of July studying
Nepenthes and their hybrids for my PhD project. I would be glad to meet
with anyone from the region or anyone who might be visiting the region
during that time to talk Nepenthes and/or explain more about my study
(Joseph, send me your phone # and I'll ring you up when I get there!).
As for Jans response to my query concerning any other
electrophoretic studies on Nepenthes: I am well aware of Tim Lowrey's
work- he is my advisor and the data you refered to is sitting on my desk
right now. Unfortunately, his study addressed whether Nepenthes show
evidence of gene duplication consistent with polyploidy (they don't) and I
am trying to study the genetic population structure in hybrid zones. Much
of his data do not have the resolution I need.
>What kind of extract are you working with? Which enzymes do you want
>to investigate? Do you use antioxidants?
Leaf extracts; any that show allelic variation between species and yes. I
used an extraction buffer originally designed for spruce that is supposed
to be great at cutting through the gunk of even the ickiest of plants (from
a protein's point of view).
>It frequently helps to add physiological concentrations of coenzymes
>of the investigated enzymes to the extraction buffer.
Nice idea, but it's a little late to be changing the extraction buffer
since I am approximately on the opposite side of the planet from my study
plants. The other problem is that while adding coenzymes to the extraction
buffer might help resolve one enzyme, they tend to screw up resolution for
other enzymes that I might potentially want to look at.
>Which detection methods do you (want to) apply? I suppose you use
>cellulose electrophoresis for your isozyme analyses.
Starch gels.
I am getting some better results now, but it is tedious experimenting trial
and error style with enzymes and buffers and I have a somewhat limited
supply of leaf extract on hand to experiment with. It would save me a lot
of time if someone else had already worked out some enzyme systems in this
region of plant taxonomy. But, if no one's done it, then I guess I get to
be the one to do the grunt work.
By the way, there is a really interesting paper on the use of insect
nutrition by carnivorous plants in last month's Oecologia (Dec 1, 1977, vol
112(4), starting on page 464). Using nitrogen stable isotope
discrimination they (I think it is Schultz et al., I don't have the paper
on my desk) were able to determine that N. mirabilis gets 67% of its
nitrogen from captured insects. They also report the figures for
Cephalotus, Darlingtonia and Drosera. Pretty neat stuff.
Chris
-----------------------------------------------------------
Chris Frazier
Dept. of Biology, UNM
Albuquerque, NM, USA 87131
(505) 277-0683
Fax: (505) 277-3781
Homepage: http://redtail.unm.edu/
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