Arab. chloroplasts

Karen.Ford 23144HEN at MSU.EDU
Fri Jun 4 10:36:00 EST 1993


Hello:  I am attempting to show chloroplast import in Arabidopsis.  At the
moment I have been successful in isolating intact chloroplasts, albeit with a
somewhat low yield.  The problem is that I am unable to show that these
chloroplasts are import competent (i.e. the precursor of RUBISCO small subunit
seems to bind, but is not processed to mature size).  I have tried to assess
whether the precursor is actually imported, and protease protected by
treatment of the intact chloroplast with thermolysin at a final concentration
of 200 ug/ml, sofar this hasn't worked (it looks like the protease is lysing
the chloroplast).

1)  Has anyone tried to import protein into isolated Arabidopsis chloroplasts?
Do you have any helpful comments on increasing yield of intact chloroplasts?

2)  Has anyone treated isolated chloroplasts with protease, and if so what
concentrations are successful?

If anyone has worked out a procedure for these experiments, they would be
greatly appreciated.  Thanks, --Erik Nielsen (a friend's account) please send
any replies to:  23144hen at ibm.cl.msu.edu



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