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Cloning of DNA of the rpoBC operon from the chromosome of Escherichia coli K12

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Summary

We provide evidence that, in terms of transcriptional organisation, the rpoBC operon carried by λrif d 18 accurately represents the corresponding region of the E. coli K12 chromosome.

A restriction fragment of E. coli K12 chromosomal DNA carrying the genes rpoBC (encoding the β and β′ subunits of RNA polymerase) and rplL (coding for ribosomal proteins L7/L12) was cloned in a λ vector, and the resulting phage tested for gene expression. In common with the corresponding fragment of λrif d 18 DNA, the chromosomal fragment has no strong promoter for rplL or rpoBC transcription. Another new phage was constructed by adding, to the restriction fragment carrying the rplL rpoBC structural genes from λrif d 18, a sequence from the E. coli K12 chromosome which includes a promoter for these genes. As in λrif d 18 itself, this promoter is shared with rplJ but not with rplKA.

The properties of the latter phage also show that the dominant rifampicin-resistance characteristic of λrif d 18 results from more than one mutation.

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References

  • Appleyard, R.K.: Segregation of new lysogenic types during growth of a doubly lysogenic strain derived from Escherichia coli K12. Genetics 39, 440–452 (1954)

    Google Scholar 

  • Austin, S.J., Tittawella, I.P.B., Hayward, R.S., Scaife, J.G.: Amber mutations of Escherichia coli RNA polymerase. Nature New Biol. 232, 133–136 (1971)

    Google Scholar 

  • Benton, W.D., Davis, R.W.: Screening λgt recombinant clones by hybridisation to single plaques in situ. Science 196, 180–182 (1977)

    Google Scholar 

  • Bolivar, F., Rodriguez, R.L., Green, P.J., Betlach, M.G., Heynecker, H.L., Boyer, H.W., Crosa, J.H., Falkow, S.: Construction and characterisation of new cloning vehicles. II. A multipurpose cloning system. Gene 2, 95–113 (1977)

    Google Scholar 

  • Boothroyd, J.C.: PhD Thesis, University of Edinburgh (1979)

  • Boyd, D., Zillig, W., Scaife, J.: Reference mutations for the β sub-unit of RNA polymerase. Mol. Gen. Genet. 130, 315–320 (1974)

    Google Scholar 

  • Fiil, N.P., Bendiak, D., Collins, J., Friesen, J.D.: Expression of Escherichia coli ribosomal protein and RNA polymerase genes cloned on plasmids. Mol. Gen. Genet. 173, 39–50 (1979)

    Google Scholar 

  • Hayward, R.S.: DNA blockade by rifamipicin-inactivated E. coli RNA polymerase, and its amelioration by a specific mutation. Eur. J. Biochem. 71, 19–24 (1976)

    Google Scholar 

  • Hughes, S.G.: A map of the cleavage sites for endonuclease AvaI in the chromosome of bacteriophage lambda. Biochem. J. 163, 503–509 (1977)

    Google Scholar 

  • Jaskunas, S.R., Lindahl, L., Nomura, M., Burgess, R.R.: Identification of two copies of the gene for the elongation factor EF-Tu in E. coli. Nature 257, 458–462 (1975)

    Google Scholar 

  • Kirschbaum, J.B., Konrad, E.B.: Isolation of a specialised lambda transducing bacteriophage carrying the beta subunit gene for Escherichia coli ribonucleic acid polymerase. J. Bacteriol. 116, 517–526 (1973)

    Google Scholar 

  • Lindahl, L., Jaskunas, S.R., Dennis, P.P., Nomura, M.: Cluster of genes in Escherichia coli for ribosomal proteins, ribosomal RNA, and RNA polymerase subunits. Proc. Natl. Acad. Sci. U.S.A. 72, 2743–2747 (1975)

    Google Scholar 

  • Lindahl, L., Yamamoto, M., Nomura, M., Kirschbaum, J.B., Allet, B., Rochaix, J.-D.: Mapping of a cluster of genes for components of the transcriptional and translational mechanisms of Escherichia coli. J. Mol. Biol. 109, 23–47 (1977)

    Google Scholar 

  • Linn, T., Scaife, J.G.: Identification of a single promoter in E. coli for rplL and rpoBC. Nature 275, 33–37 (1978)

    Google Scholar 

  • Maniatis, T., Jeffrey, A., van de Sande, H.: Chain length determination of small double- and single-stranded DNA molecules by polyacrylamide gel electrophoresis. Biochemistry 14, 3787–3794 (1975)

    Google Scholar 

  • MeDonell, M.W., Simon, M.N., Studier, F.W.: Analysis of restriction fragments of T7 DNA and determination of molecular weights by electrophoresis in neutral and alkaline gels. J. Mol. Biol. 110, 119–146 (1977)

    Google Scholar 

  • Nakamura, Y., Yura, T.: Effects of rifampicin on synthesis and functional activity of DNA-dependent RNA polymerase in Escherichia coli. Mol. Gen. Genet. 145, 227–237 (1976)

    Google Scholar 

  • Newman, A.J., Linn, T.G., Hayward, R.S.: Evidence for co-transcription of the RNA polymerase genes rpoBC with a ribosomal protein gene of Escherichia coli. Mol. Gen. Genet. 169, 195–204 (1979)

    Google Scholar 

  • Post, L.E., Strycharz, G.D., Nomura, M., Lewis, H., Dennis, P.P.: Nucleotide sequence of the ribosomal protein gene cluster adjacent to the gene for RNA polymerase subunit β in Escherichia coli. Proc. Natl. Acad. Sci. U.S.A. 76, 1697–1701 (1979)

    Google Scholar 

  • Rigby, P.W.J., Diekmann, M., Rhodes, C., Berg, P.: Labelling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J. Mol. Biol. 113, 237–251 (1977)

    Google Scholar 

  • Rossi, J.J., Ross, W., Egan, J., Lipman, D.J., Landy, A.: Structural organisation of Escherichia coli tRNATyr gene clusters in four different transducing bacteriophage. J. Mol. Biol. 128, 21–47 (1979)

    Google Scholar 

  • Schweitzer, S.M., Matzura, H.: Transformation of Escherichia coli by a specific DNA restriction fragment. Mol. Gen. Genet. 155, 213–217 (1977)

    Google Scholar 

  • Shimada, K., Weisberg, R.A., Gottesman, M.E.: Prophage lambda at unusual chromosomal locations. II. The components of the secondary attachment sites. J. Mol. Biol. 93, 415–429 (1975)

    Google Scholar 

  • Sutcliffe, J.G.: pBR322 restriction map derived from the DNA sequence: accurate DNA size markers up to 4361 nucleotide pairs long. Nucleic Acids Res. 5, 2721–2728 (1978)

    Google Scholar 

  • Yamamoto, M., Nomura, M.: Co-transcription of genes for RNA polymerase sub-units β and β′ with genes for ribosomal proteins in Escherichia coli. Proc. Natl. Acad. Sci. U.S.A. 75, 3891–3895 (1978)

    Google Scholar 

  • Yamamoto, M., Nomura, M.: Organisation of genes for transcription and translation in the rif region of the Escherichia coli chromosome. J. Bacteriol. 137, 584–594 (1979)

    Google Scholar 

  • Zissler, J., Signer, E., Schaefer, F.: The role of recombination in growth of phage λ II. Inhibition of growth by prophage P2. In: The bacteriophage lambda (A.D. Hershey, ed.). New York: Cold Spring Harbor Laboratory 1971

    Google Scholar 

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Communicated by A. Böck

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Newman, A., Hayward, R.S. Cloning of DNA of the rpoBC operon from the chromosome of Escherichia coli K12. Molec. Gen. Genet. 177, 527–533 (1980). https://doi.org/10.1007/BF00271493

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  • DOI: https://doi.org/10.1007/BF00271493

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