Isolation of ribonuclease-free polysomes from human placenta.
Human placenta is known to have a high level of polysome-bound ribonuclease which has hindered the isolation of intact polyribosomes from this tissue. We describe conditions for preparing polysomes devoid of apparent ribonuclease activity from both first trimester and term placenta. This stable preparation was achieved by utilizing buffer at low pH containing 300 mM LiCl and precipitating the polysomes chemically with 200 mM MgCl2. The yield of polysomes obtained by this procedure is 2-2.5 fold greater than that obtained by the conventional method of preparing placental polysomes. The polysomes are considered pure as judged by the ratios of absorbance 260/280 and 260/235. Moreover, the yield of polysomes obtained is greater than 95% of the tissue content and the profile of the polysomes is probably representative of the in vivo population. This is concluded from experiments in which a known amount of labelled chick polysomes was added to fresh placental tissue and the recovery of label and its distribution was analyzed.
D002458:Cell Fractionation; D002499:Centrifugation, Density Gradient; D005260:Female; D006801:Humans; D008970:Molecular Weight; D010920:Placenta / Q000648:ultrastructure*; D011132:Polyribosomes / Q000032:analysis*; D011247:Pregnancy; D011261:Pregnancy Trimester, First; D011263:Pregnancy Trimester, Third; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000302:isolation & purification; D012260:Ribonucleases / Q000378:metabolism; D013056:Spectrophotometry, Ultraviolet
D012333:RNA, Messenger; D012260:Ribonucleases