Abstract
An original method to isolate nuclei from the posterior part of the silk glands has been developed. After a collagenase and Triton X-100 treatment, silk glands were filtered through a steel sieve. This step, which is the most efficient one for the purification, is followed by several washings. The preparation of nuclei is fairly pure, RNA: DNA ratio being 0.3 at the end of the whole procedure, and the final DNA recovery quite satisfactory (40-60%). Although chromatin of the purified nuclei is unusually condensed, and in spite of RNase activity, RNA transcription measured in vitro is quantitatively significant (0.01 % of the total DNA). This transcription, resulting from the activity of endogenous RNA polymerases, reaches a maximum when nuclei were extracted from animals on the 4th day of the fifth instar. Differences with results obtained in vivo are discussed.
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