In vitro transcription of a transfer RNA gene.
The Escherichia coli tRNA(Tyr) gene carried by the varphi80psu(3) + transducing phage was transcribed in vitro by DNA-dependent RNA polymerase. The enzymatically synthesized tRNA(Tyr)-like polynucleotide chains were detected by competition with purified E. coli(32)P-tRNA(Tyr) for specific hybridization sites on varphi80psu(3) + DNA. Analysis by sucrose gradient centrifugation showed the tRNA(Tyr)-like chains to possess a heterogeneous distribution with respect to sedimentation coefficients, with a broad peak around 8 S. The presence of the termination factor rho during the transcription did not significantly reduce the average size of the in vitro synthesized tRNA(Tyr)-like chains.
D001435:Bacteriophages / Q000378:metabolism; D002247:Carbon Isotopes; D002499:Centrifugation, Density Gradient; D002848:Chromatography, DEAE-Cellulose; D004247:DNA / Q000302:isolation & purification; D004926:Escherichia coli / Q000201:enzymology / Q000378:metabolism*; D005815:Genetic Code; D006824:Hybridization, Genetic; D066298:In Vitro Techniques; D010759:Phosphorus Isotopes; D012313:RNA / Q000096:biosynthesis; D012316:RNA Nucleotidyltransferases / Q000378:metabolism; D012343:RNA, Transfer / Q000378:metabolism; D014316:Tritium
D002247:Carbon Isotopes; D010759:Phosphorus Isotopes; D014316:Tritium; D012313:RNA; D004247:DNA; D012343:RNA, Transfer; D012316:RNA Nucleotidyltransferases