Synthesis of DNAs complementary to human ribosomal RNAs polyadenylated in vitro.
Conditions are described under which poly(A) polymerase from Escherichia coli ribosomes will catalyse the addition of AMP residues onto the 3'-ends of human 18 S and 28 S ribosomal RNAs at an average rate of 40 AMP residues per 1000 nucleotides in 20 min. Single-stranded complementary DNAs (cDNAs) can then be transcribed from the polyadenylated RNAs with RNA-directed DNA polymerase from avian myeloblastosis virus. All of the sequences in the RNAs are represented in the cDNAs; measurements of the rates at which the cDNAs hybridize with their template RNAs showed that, when appropriate adjustments for differences in lengths and G + C contents of the reacting sequences are taken into account, the Rot 1/2 values of homologous RNA-cDNA hybridization reactions are directly proportional to the base-sequence complexity of the RNAs.
D004247:DNA / Q000096:biosynthesis*; D004926:Escherichia coli / Q000201:enzymology; D006801:Humans; D007700:Kinetics; D008970:Molecular Weight; D009691:Nucleic Acid Denaturation; D009693:Nucleic Acid Hybridization; D011061:Poly A / Q000096:biosynthesis*; D011062:Polynucleotide Adenylyltransferase / Q000378:metabolism; D012335:RNA, Ribosomal / Q000378:metabolism*; D014158:Transcription, Genetic
D012335:RNA, Ribosomal; D011061:Poly A; D004247:DNA; D011062:Polynucleotide Adenylyltransferase