Cell-free protein synthesis in t,e rabbit liver ribosomal system. II. Large scale preparation of purified 80S ribosomes.
A procedure for the preparation of a large quantity of biologically active, highly purified ribosomes from rabbit liver is described. The method employs polyethylene glycol-dextran sulfate parition and DEAE-cellulose chromatography to overcome the limitations encountered in conventional procedures. The entire process takes only 48 h to obtain 10,000 A(260) units of ribosomes. The ribosomes thus obtained are predominantly 78S particles with a constant protein-RNA ratio of 0.95. The ribosomes are free from RNase, amino-acyl-tRNA synthetase, and amino-acyl-tRNA: protein transferase activity. The protein synthesizing activity is dependent on added mRNA and protein factors. These ribosomes are stable for prolonged periods of storage in a liquid nitrogen refrigerator.
D000604:Amino Acyl-tRNA Synthetases / Q000032:analysis; D000818:Animals; D002458:Cell Fractionation; D002474:Cell-Free System; D008099:Liver / Q000378:metabolism*; D010649:Phenylalanine / Q000378:metabolism; D011119:Polynucleotides / Q000378:metabolism; D012343:RNA, Transfer; D011817:Rabbits; D012156:Reticulocytes / Q000201:enzymology; D012260:Ribonucleases / Q000032:analysis; D012269:Ribosomal Proteins / Q000096:biosynthesis*; D012270:Ribosomes / Q000378:metabolism; D014500:Uracil Nucleotides / Q000378:metabolism
D011119:Polynucleotides; D012269:Ribosomal Proteins; D014500:Uracil Nucleotides; D010649:Phenylalanine; D012343:RNA, Transfer; D012260:Ribonucleases; D000604:Amino Acyl-tRNA Synthetases