title: The nucleotide sequence in the promoter region of the fene for an Escherichia coli tyrosine transfer ribonucleic acid.
abstract: The sequence of 29 nucleotides in the region preceding the initiation of the transcription of the Escherichia coli tyrosine tRNA gene has been determined. This is: (5')  ---GGGGCGCATCATATCAAATGACGCGCCGC--- (3') (3') ---CCCCGCGTAGTATAGTTTACTGCGGCGGCG--- (5'). The general approach used for the sequence determination involved the DNA polymerase I-catalyzed elongation of suitable deoxyribopolynucleotide primers when hybridized to the 1-strand of phi80psuIII plus DNA at the appropriate site. Sequences of the newly grown oligonucleotide chains were determined by a combination of two-dimensional fingerprinting following partial exonucleolytic degradation, nearst neighbor analyses, and determination of pyrimidine tracts. Primer elongations were carried out in a controlled and stepwise manner and the newly grown oligonucleotide chains were kept short by incorporating the following features into the method: (a) the insertion of a ribonucleotide unit at or near the 3' terminus of the primers; (b) the use of a maximum of three nucleoside 5'-triphosphates in the first stage of the elongation reaction isolation of the elongated primer, and its reuse in a second step together with different sets of deoxy-nucleoside triphosphates; and (c) elongation of the primer using all of the four nucleoside triphosphates with one of the triphosphates being supplied in a limiting concentration.
mesh_terms: D001483:Base Sequence; D002855:Chromatography, Thin Layer; D004254:DNA Nucleotidyltransferases; D004269:DNA, Bacterial / Q000378:metabolism; D004926:Escherichia coli / Q000032:analysis* / Q000378:metabolism; D005796:Genes; D007700:Kinetics; D009693:Nucleic Acid Hybridization; D009713:Nucleotidyltransferases; D009841:Oligonucleotides / Q000032:analysis; D011118:Polynucleotide Ligases; D012329:RNA, Bacterial / Q000096:biosynthesis*; D012343:RNA, Transfer / Q000096:biosynthesis*; D012265:Ribonucleotides / Q000032:analysis; D014158:Transcription, Genetic; D014443:Tyrosine
chemical_list: D004269:DNA, Bacterial; D009841:Oligonucleotides; D012329:RNA, Bacterial; D012265:Ribonucleotides; D014443:Tyrosine; D012343:RNA, Transfer; D004254:DNA Nucleotidyltransferases; D009713:Nucleotidyltransferases; D011118:Polynucleotide Ligases