Isolation of RNA with properties of messenger RNA from cerebral polyribosomes.
RNA which dissociated from purified cerebral polyribosomes of adult rats in the presence of EDTA was isolated by fractionation in a discontinuous sucrose gradient. The yield was 2% of the total polyribosomal RNA. The base composition resembled the complementary values for rat DNA and was very different from base compositions of ribosomal RNA and transfer RNA. This RNA fraction contained a large proportion of molecules which were rapidly labeled in vivo and hybridized to homologous DNA. The polyribosomal RNA preparation also exhibited high template activity in a cerebral cell-free system which had previously been stripped of the capacity to incorporate amino acids in the absence of added messenger RNA (mRNA). Sedimentation analysis revealed only two peaks, with coefficients of approximately 8 S and 16 S. The data indicate that RNA with the properties of mRNA can be selectively isolated from cerebral polyribosomes under mild conditions which avoid degradation.
D000818:Animals; D001483:Base Sequence; D002247:Carbon Isotopes; D002474:Cell-Free System; D002499:Centrifugation, Density Gradient; D002540:Cerebral Cortex / Q000032:analysis*; D006863:Hydrogen-Ion Concentration; D008297:Male; D009693:Nucleic Acid Hybridization; D010649:Phenylalanine / Q000378:metabolism; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000302:isolation & purification*; D012335:RNA, Ribosomal / Q000302:isolation & purification; D012343:RNA, Transfer / Q000302:isolation & purification; D051381:Rats; D012270:Ribosomes / Q000032:analysis*; D013395:Sucrose; D013698:Templates, Genetic
D002247:Carbon Isotopes; D012333:RNA, Messenger; D012335:RNA, Ribosomal; D010649:Phenylalanine; D013395:Sucrose; D012343:RNA, Transfer