The Journal of Cell Biology, Vol 87, 292-296, Copyright © 1980 by The Rockefeller University Press
Isolation of giant silk fibroin polysomes and fibroin mRNP particles using a novel ribonuclease inhibitor, hydroxystilbamidine
PM Lizardi
Hydroxystilbamidine isethionate, a dye capable of binding to both DNA and
RNA, has been found to be a powerful inhibitor of cellular ribonucleases. A
procedure has been developed that, with the aid of this compound, permits
the preparative isolation of giant silk fibroin polyribosomes from the
posterior silk gland of Bombyx mori. The polyribosomes contain
approximately 45-112 ribosomal particles, as judged by electron microscopy.
Treatment of giant fibroin polyribosomes with EDTA releases a particle that
sediments at 125S. This mRNP particle contains biologically active silk
fibroin mRNA, as judged by cell-free translation in an mRNA-dependent
reticulocyte cell-free system.