multiple sequences from Edman degrad

Bilbrey Robert C478SCB at SEMOVM.SEMO.EDU
Tue Oct 24 18:49:09 EST 1995


Hi Netters,

I had a brain spasm regarding generating multiple sets of sequence
data from a series of manual Edman degradations, and wanted to see
if anyone would be good enough to tell me why this can't work.

I want to purify some intracellular fungal proteases and determine
enough sequence data to design degenerate probes. I have the purif.
worked out and am going to do the sequencing manually. It seems to
me that I should be able to determine 10-12 N-termnal residues before
the background gets to be a problem. Since the activities that I am
interested in are a royal pain to isolate, I'd like to cleave the
bulk of the remaining polypeptide with CNBr, purify the fragments
and get some internal sequence data from one or more of these pieces.

Think it will work? Any comments wil be greatly appreciated, especially
as I've never done any sequencing and can't afford to send the samples
off for out of house sequencing.

Robert Bilbrey
c478scb at semovm.semo.edu





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