Protein composition of lysin fractions
One ml of each fraction was extracted with 250 microliters of phenol. Two aliquots of samples 9 and 10 were phenol extracted, one with 4 microliters of 6mg/ml ovalbumin added and one with no addtions. Proteins were precipitated with 0.7ml acetone (kept 45' at -20C). Pellets were washed once with acetone. Samples 9 and 10 were dissolved in 75 microliters of cracking buffer and. Samples 13 and 14 were dissolved in 35 microliters of cracking buffer. Samples were reduced and alklyated and then divided into two fractions one of which was dansylated and the other of which was applied to the gel w/o dansylation. 12 microliter aliquots were applied to the gel which was photographed for fluorescence and then stained with Coomassie Brilliant Blue.
Samples 9 and 10 w/o added ovalbumin contained obvious precipitate upon acetone treatment and adding ovalbumin did not notably increase precipitate volume. Note that direct staining with Coomassie Blue gives sharper bands. Samples 13 and 14 appeared to give less precipitate. Note the material at the top of the gel with samples 13 and 14 which stains clearly with dansyl chloride, but not with Coomassie blue.
