could protein from dried gel be sequenced?

J. Norrie 4104jnor at vm1.ulaval.ca
Wed May 24 10:02:20 EST 1995


Hello bionetters,  
 I have purified protein of interest and have done few biochemical
characterisations. The problem is that this protein is induced in young
leaves in response to chemical stimulus. A one step purification protocol
enables me to isolate the protein, but the quantity of protein of interest
is tooooo low. I have dried some gels with my pure(fairly..80%) protein. I
want to sequence this protein in order to fish out the gene of interest,
for which I want to use either immunoscreening or would like to prepare
degenerate oligomers and do a PCR on the polyA to get the cDNA. 
Is there a method to isolate the protein from fixed coummassie stained
dried SDS-PAGE? so as to be used for protein sequencing? and are there
major advantages and disadvantages of this method?
Thank you,  
Savita
Centre for research in Horticulture,   
Pav. Envirotron,
Univ. Laval, Quebec City   
Email: 4104vsav at vm1.ulaval.ca



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