RIbosome isolation

Kyle Legate legatek at mcmail.cis.mcmaster.ca
Tue Dec 17 09:03:47 EST 2002


On 16 Dec 2002 oustudygirl at hotmail.com wrote:

> I need to isolate whole, intact and functional ribosomes from plant leaf =
tissue, while diminishing RuBisco or removing chloroplast. All the methods =
I have found were either to isolate whole mammalian ribosomes, or isolate r=
ibosomal subunits in plants. I`ve been trying to combine and create an adeq=
uate method but I get very low yeilds or RuBisco contamination. Can any hel=
p=C9
>
What did you try? I think that once you obtain a ribosome-containing
supernatant after a series of spins to remove cellular debris and
chloroplasts, the supernatant can be spun over a 10-40% sucrose gradient,
and the ribosomes collected by bottom puncture and monitoring at A260 nm.
I know RuBisco's pretty big, but I don't think it would comigrate with
ribosomes in a sucrose gradient--does it?

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legatek at mcmaster.ca=09=09Kyle Legate            legatek at hotmail.com

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