Skip to main content
SpringerLink
Log in

Use of Mu phages to isolate transposon insertions juxtaposed to given genes ofEscherichia coli

  • Published:
Current Microbiology Aims and scope Submit manuscript

Abstract

The small sizes of the DNA fragments transduced by lysates of phage Mu and of mixed lysates of Mu and mini-Mu18A-1 (an internally deleted Mu phage) provide a method for the selection of insertions of transposon Tn10 located very close to givenEscherichia coli genes. Generalized transduction with Mu lysates selected for those insertions located within 38 kilobase pairs of the gene of interest whereas insertions located within about half that distance are directly selected by use of mini-Mu phages. Use of these transduction systems avoids screening of individual colonies by phage P1 transduction for those transposon insertions closely linked to a given gene. Such insertions are most useful for localized mutagenesis and for in vitro molecular cloning.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Literature Cited

  1. Bachmann, B. J., Low, K. B. 1980. Linkage map ofEscherichia coli K-12, edition 6. Microbiological Review44:1–56.

    PubMed  Google Scholar 

  2. Bachmann, B. J., Low, K. B., Taylor, A. L. 1976. Recalibrated linkage map ofEscherichia coli K-12. Bacteriological Review40:116–167.

    Google Scholar 

  3. Bode, E. G., Howe, M. M., Rawluk, L. 1978. Preferential generalized transduction by bacteriophage Mu. Molecular and General Genetics160:89–94.

    Google Scholar 

  4. Bochner, B. R., Huang, H., Schieven, G. L., Ames, B. N. 1980. Positive selection for loss of tetracycline resistance. Journal of Bacteriology143:926–933.

    PubMed  Google Scholar 

  5. Chang, Y-Y., Cronan, Jr., J. E. 1982. Mapping nonselectable genes ofEscherichia coli by using transposon Tn10: location of a gene affecting pyruvate oxidase. Journal of Bacteriology151:1279–1289.

    PubMed  Google Scholar 

  6. Clarke, L., Carbon, J. 1976. A colony bank containing synthetic Col El hybrid plasmids representative of the entireE. coli genome. Cell9:91–99.

    PubMed  Google Scholar 

  7. Cobbett, C. S., Pittard, J. 1980. Formation of a λ(Tn10)tyrR + specialized transducing bacteriophage fromEscherichia coli K-12. Journal of Bacteriology144:877–883.

    PubMed  Google Scholar 

  8. Cronan, Jr., J. E., Littel, K. E., Jackowski, S. 1982. Genetic and biochemical analyses of pantothenate biosynthesis inEscherichia coli andSalmonella typhimurium. Journal of Bacteriology149:916–922.

    PubMed  Google Scholar 

  9. Cronan, Jr., J. E., Silbert, D. F., Wulff, D. L. 1972. Mapping of thefabA locus for unsaturated fatty acid biosynthesis inEscherichia coli. Journal of Bacteriology112:206–211.

    PubMed  Google Scholar 

  10. Desmet, R. L., Faelen, M., Lefebvre, N., Resibois, A., Toussaint, A., von Gijsegem, F. 1980. Genetic study of Mu transposition and Mu-mediated chromosomal rearrangements. Cold Spring Harbor Symposium of Quantitative Biology45:355–363.

    Google Scholar 

  11. Faelen, M., Toussaint, A., Resibois, A. 1979. Mini-Mu-duction: a new mode of gene transfer mediated by mini-Mu. Molecular and General Genetics176:191–197.

    PubMed  Google Scholar 

  12. Gill, G. S., Hull, R. C., Curtiss, III, R., 1981. Mutator bacteriophage D108 and its DNA: an electron microscopic characterization. Journal of Virology37:420–430.

    PubMed  Google Scholar 

  13. Howe, M. M. 1973. Transduction by bacteriophage Mu-1. Virology55:103–117.

    PubMed  Google Scholar 

  14. Kleckner, N. 1981. Transposable elements in prokaryotes. Annual Review of Genetics15:341–404.

    PubMed  Google Scholar 

  15. Kleckner, N., Roth, J., Botstein, D. 1977. Genetic engineeringin vivo using translocatable drug-resistance elements: new methods in bacterial genetics. Journal of Molecular Biology116:125–159.

    PubMed  Google Scholar 

  16. Leach, D., Symonds, N. 1979. The isolation and characterization of a plaque-forming derivative of bacteriophage Mu carrying a fragment of Tn3 conferring ampicillin resistance. Molecular and General Genetics172:179–184.

    PubMed  Google Scholar 

  17. Lefebvre, N., Toussaint, A. 1981. Transfer ofSalmonella typhimurium andKlebsiella pneumoniae genes inE. coli K12 by mini-Mu-duction. Molecular and General Genetics181:268–272.

    PubMed  Google Scholar 

  18. Maloy, S. R., Nunn, W. D. 1981. Selection for loss of tetracycline resistance byEscherichia coli. Journal of Bacteriology145:1110–1112.

    Google Scholar 

  19. Mise, K. 1971. Isolation and characterization of a new generalized transducing bacteriophage different from P1 inEscherichia coli. Journal of Virology7:168–175.

    PubMed  Google Scholar 

  20. Resibois A., Toussaint, A., Van Gijsegem, F., Faelen, M. 1981. Physical characterization of mini-Mu and mini-D108 derivatives. Gene14:103–113.

    PubMed  Google Scholar 

  21. Susskind, M. M., Botstein, D. 1978. Molecular genetics of bacteriophage P22. Microbiological Review42:385–413.

    Google Scholar 

  22. Toussaint, A., Faelen, M., Resibois, A. 1981. Chromosomal rearrangements induced by mini-Mu and mini-D108: mini review and new data. Gene14:115–119.

    PubMed  Google Scholar 

  23. Vogel, H. J., Bonner, D. M. 1956. Acetyl-ornithinase ofEscherichia coli: partial purification and some properties. Journal of Biological Chemistry218:97–116.

    PubMed  Google Scholar 

  24. Waggoner, B., Pato, M., Toussaint, A., Faelen, M. 1981. Replication of mini-Mu prophage DNA. Virology113:379–387.

    PubMed  Google Scholar 

  25. Wu, T. T. 1966. A model for three point analyses of random general transduction. Genetics54:405–410.

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Microbiology, University of Illinois, 61801, Urbana, Illinois, USA

    John E. Cronan Jr.

Authors
  1. John E. Cronan Jr.
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Cronan, J.E. Use of Mu phages to isolate transposon insertions juxtaposed to given genes ofEscherichia coli . Current Microbiology 9, 245–252 (1983). https://doi.org/10.1007/BF01567195

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01567195

Keywords

Search

Navigation