immunolocalization collaboration

Paul Roessler paulr at nrel.nrel.gov
Tue Jan 18 13:52:11 EST 1994




Wanted:  Collaborator

Is there anyone out there who would be interested in a
collaborative effort to determine the cellular location of a
multifunctional type of acetyl-CoA carboxylase ("ACCase", an
important enzyme for fatty acid synthesis and elongation) in the
diatom Cyclotella cryptica?  Based on the deduced amino acid
sequence, the polypeptide appears to have an ER signal sequence,
and thus may enter into the chloroplast (where de novo fatty acid
synthesis typically occurs) after first going through the ER (see
Gibbs, S.P. (1979) J. Cell Sci. 35:253-266; and Bhaya, D. and
Grossman, A. (1991) Mol. & Gen. Genet. 229:400-404). However,
diatoms make substantial quantities of very long chain fatty
acids, and since elongation of chloroplast-produced fatty acids
occurs outside of the chloroplast (probably within the ER), there
may be a need for ACCase in this compartment, also.  Basically,
I'm interested in knowing whether the ACCase that I've purified
(and cloned) is found in the chloroplast, the ER, or both.

I have very specific, affinity-purified polyclonal antibodies
already available.  What I need is someone who is experienced in
immunolocalization techniques, since my knowledge in this area is
nearly nonexistent.

Thanks, Paul Roessler
        National Renewable Energy Lab
        Golden, Colorado

        paulr at nrel.gov






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