Summary
This paper reports a cleavage site map of Tn5 for restriction enzymes BamHI, BglI, BglII, HindII, HindIII, HpaI, SalI, AvaI, SmaI, XhoI, PstI, PvuII, HaeII and HaeIII that was determined by the analysis of restriction enzyme cleavage patterns of ColEl, two independent ColE1::Tn5 plasmids, and a ColE1::Tn5 deletion derivative. BalI, EcoRI, KpnI, and PvuI do not cleave Tn5. Construction and analysis of in vitro-generated deletions of a ColE1::Tn5 plasmid limit the sequences encoding neomycin resistance to a 1500-base-pair-long segment of Tn5. Insertion of DNA at a BglII site within this segment results in loss of the neomycin resistance phenotype. Since this BglII site lies in an inverted repeat region, sequences within this repeat seem to be involved in the expression of neomycin resistance.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bachmann, B., Bachmann, J.: Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol. Rev. 36, 525–557 (1972)
Barnes, W.: Plasmid detection and sizing in single colony lysates. Sci. 195, 363–394 (1977)
Berg, D., Davies, J., Allet, B., Rochaix, J.D.: Transposition of R-factor genes to bacteriophage λ. Proc. Natl. Acad. Sci. U.S.A. 72, 3628–3632 (1975)
Berg, D., Jorgensen, R., Davies, J.: Transposable Kanamycin-Neomycin resistance determinants. In: Microbiology-1978, D. Schessinger (ed.)., pp. 13–15, Washington, D.C.: ASM Publications 1978
Cameron, J., Philippsen, P., Davis, R.: Analysis of chromosomal integration and deletions of yeast plasmids. Nucleic Acids Res. 4, 1429–1448 (1977)
Collins, J., Johnsen, M., Jorgensen, P., Valentin-Hansen, P., Karlström, H., Gautier, F., Lindenmaier, W., Mayer, H., Sjöberg, B.: Expression of plasmid genes: ColE1 and derivatives. In: Microbiology-1978, D. Schlessinger (ed.), pp. 150–153. Washington, D.C.: ASM Publications 1978
Heffron, F., Sublett, R., Hedges, R., Jacob, A., Falkow, S.: Origin of the TEM beta-lactamase gene found on plasmids. J. Bacteriol. 122, 250–256 (1975)
Humphreys, S.O., Willshaw, G.A., Anderson, E.S.: A simple method for the preparation of large quantities of pure plasmid DNA. Biochim. Biophys. Acta 383, 457–463 (1975)
Kleckner, N., Chan, R., Tye, B.K., Botstein, D.: Mutagenesis by insertion of a drug-resistance element carrying an inverted repetition. J. Mol. Biol. 97, 561–575 (1975)
Mandel, M., Higa, A.: Calcium-dependent bacteriophage DNA infection. J. Mol. Biol. 43, 159–162 (1970)
Maniatis, T., Jeffrey, A., vandde Sande, H.: Chain length determination of small double- and single-stranded DNA molecules by polyacrylamide gel electrophoresis. Biochemistry 14, 3787–3794 (1975)
Meagher, R., Tait, R., Betlach, M., Boyer, H.: Protein expression in E. coli minicells by recombinant plasmids. Cell 10, 521–536 (1977)
Oka, A.: Fine cleavage map of a small colicin E1 plasmid carrying genes responsible for replication and colicin E1 immunity. J. Bacteriol. 133, 916–924 (1978)
Oka, A., Takanami, M.: Cleavage map of colicin E1 plasmid. Nature 264, 193–196 (1976)
Postle, K., Reznikoff, W.S.: Identification of the Escherichia coli tonB gene product in minicells containing tonB hybrid plasmids. J. Mol. Biol. 131, 619–636 (1979)
Shinnick, T.M., Lund, E., Smithies, O., Blattner, F.R.: Hybridization of labelled RNA to DNA in agarose gels. Nucleic Acids Res. 2, 1911–1929 (1975)
Smith, H.O., Wolcox, K.W.J.: A restriction enzyme from Haemophilus influenzae. I. Purification and general properties. J. Mol. Biol. 51, 376–391 (1970)
Author information
Authors and Affiliations
Additional information
Communicated by A.J.Clark
Rights and permissions
About this article
Cite this article
Jorgensen, R.A., Rothstein, S.J. & Reznikoff, W.S. A restriction enzyme cleavage map of Tn5 and location of a region encoding neomycin resistance. Molec. Gen. Genet. 177, 65–72 (1979). https://doi.org/10.1007/BF00267254
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00267254