The nucleotide sequence in the promoter region of the gene for an Escherichia coli tyrosine transfer ribonucleic acid

Journal of Biological Chemistry (Impact Factor: 4.57). 09/1976; 251(15):4481-9.
Source: PubMed


The sequence of the first 29 nucleotides in the promoter region of a tyrosine tRNA gene has previously been determined (Sekiya, T., van Ormondt, H., and Khorana, H.G. (1975) J. Biol. Chem. 250, 1087-1098). This work has now been extended to give the sequence of a total of 59 nucleotides; the sequence is as follows: (see article). The general approach used in the determination of the sequence involved the DNA polymerase I-catalyzed elongation of synthetic deoxyribopolynucleotide primers hydridized to the l-strand of phi80psu+III DNA at the appropriate site. Sequencing of the newly added nucleotides was facilitated by the use of a number of techniques including (a) elongation of the primer with the use of all of the four nucleoside 5'-triphosphates but limiting the concentration of one of the triphosphates, (b) insertion of ribonucleotide units at appropriate sites so as to permit subsequent specific cleavages by pancreatic RNase, and (c) two-dimensional fingerprinting of the oligonucleotides in conjunction with partial exonucleolytic degradation, comprehensive nearest neighbor analyses, and the determination of pyrimidine tracts.

Full-text preview

Available from:
  • [Show abstract] [Hide abstract]
    ABSTRACT: Restriction endonucleases EcoRI and HindIII generated fragments of T4 cytosine-containing DNA were inserted into bacteriophage vector lambdagtSuIII and plasmid vectors pMB9 and pBR313. Resulting clones were screened for hybridization with 32P labeled T4 tRNA. Recombinant bacteriophages and plasmids were isolated which contained a T4 fragment coding for T4 RNA species 1 and 2 and T4 tRNA Arg. Selected lambda-T4 hybrid bacteriophages were grown to high titer and their DNA analyzed by gel electrophoresis.
    No preview · Article · Feb 1976 · Gene
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: We have derived the nucleotide sequence of a segment of the operator-promoter region of the galactose operon of E. coli, by using a variety of DNA sequencing analyses. We have previously reported the sequence of the 5' terminal portion of gal mRNA [Musso, R. E., de Crombrugghe, B., Pastan, I., Sklar, J., Yot, P. & Weissman, S. (1974) Proc. Natl. Acad. Sci. USA 71, 4940-4944] and of the 59 base pairs preceding the startpoint of gal transcription (J. Sklar, S. Weissman, R. Musso, R. Di Lauro, & B. de Crombrugghe, submitted). In conjunction with those results, the present data provide the sequence of the gal operator-promoter region. This sequence is compared with similar sequences in other promoters and operators. Tentative mechanisms for the regulation of the galactose operon are discussed.
    Full-text · Article · Feb 1977 · Proceedings of the National Academy of Sciences
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: The promoter for the major coat protein gene of bacteriophage fd contains a unique sequence, TATAAT, in the non-transcribed region corresponding to the Pribnow box. A R.Hha I cleavage site which destroys promoter function is located five base pairs upstream from the TATAAT sequence(fifteen base pairs upstream from the RNA initiation site). The promoter was cleaved into two fragments by R.Hha I and each promoter fragment was joined to DNA fragments derived from other regions. Ligation of the TATAAT-containing fragment to any of the DNA fragments examined resulted in recovery of promoter function. The results suggest for this type of promoter that no unique sequence is necessary upstream from the R.Hha I cleavage site although a contiguous DNA chain must be present in this area.
    Preview · Article · Aug 1977 · Nucleic Acids Research
Show more