Purification of polysomes.
Isolated polysomes (also known as translating ribosomes or polyribosomes) are mRNA-ribosome complexes that are frequently used for the in vitro study of the regulation of protein synthesis. Here we describe a protocol for the isolation of prokaryotic and plant polysomes by sucrose gradient sedimentation. The protocol allows for the separation of multiple ribosomes attached to mRNA from run-off ribosome monomers. In addition to collecting the polysome pellet, if the sucrose gradients used in this protocol are fractionated, it is possible to isolate and quantify the polysomes, the ribosome monomers, and the ribosomal subunits.
Department of Biology and Center for the Study of Biological Complexity, Virginia Commonwealth University, Richmond, Virginia 23284;
Maria C
MC
Rivera
Primary Pharmacology Group, Pfizer Global Research and Development, Groton, Connecticut 06340;
Bruce
B
Maguire
Department of Molecular, Cell and Developmental Biology and Department of Human Genetics, University of California, Los Angeles, California 90095.
James A
JA
Lake
D002499:Centrifugation, Density Gradient / Q000379:methods*; D059828:Plant Cells; D011132:Polyribosomes; D011387:Prokaryotic Cells; D012333:RNA, Messenger / Q000302:isolation & purification*
D012333:RNA, Messenger