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Restriction and modification in B. subtilis

Purification and general properties of a restriction endonuclease from strain R

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Summary

All Bacillus subtilis R-type strains showing the phenomena of restriction and modification contain an endonuclease that inactivates in vitro the biological activity of a variety of DNAs lacking R-specific modification, such as transfecting SPP1, SPO2 and ϕ105 DNA, and transforming B. subtilis 168-type DNA. The corresponding DNAs carrying R-specific modification are resistant to the enzyme. The enzyme has been purified approximately 400-fold and is essentially free from contaminating double strand-directed unspecific exo-or endonuclease activity. Only Mg2+ is required as cofactor. The substrate DNAs are cleaved at specific sites. The double-stranded fragments produced from SPP1 DNA (molecular weight 2.5×107) have an average molecular weight of about 3×105.

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References

  • Arber, W.: DNA modification and restriction. Prog. Nucleic Acid Res. Mol. Biol. 14, 1–37, Acad. Press Inc. (1974)

    Google Scholar 

  • Arber, W., Linn, S.: DNA modification and restriction. Ann. Rev. Biochem. 38, 467–500 (1969)

    Google Scholar 

  • Benzinger, R.: Restriction of infectious bacteriophage fd DNA's and an assay for in vitro host-controlled restriction and modification. Proc. nat. Acad. Sci. (Wash.) 59, 1294–1299 (1968)

    Google Scholar 

  • Bigger, C.H., Murray, K., Murray, N.E.: Recognition sequence of a restriction enzyme. Nature (Lond.) New Biol. 244, 7–10 (1973)

    Google Scholar 

  • Biswal, N., Kleinschmidt, A.K., Spatz, H.Ch., Trautner, T.A.: Physical properties of the DNA of bacteriophage SP50. Molec. gen. Genet. 100, 39–55 (1967)

    Google Scholar 

  • Boyer, H.W.: DNA restriction and modification mechanisms in bacteria. Ann. Rev. Microbiol. 25, 153–176 (1971)

    Google Scholar 

  • Bron, S., Murray, K.: Restriction and modification in B. subtilis. Nucleotide sequence recoginsed by restriction endonuclease R. Bsu R from strain R. Molec. gen. Genet 143, 25–33 (1975)

    Google Scholar 

  • Bron, S., Venema, G.: Ultraviolet inactivation and excision-repair in Bacillus subtilis. I. Construction and characterization of a transformable eightfold auxotrophic strain and two ultravioletsensitive derivatives. Mutation Res. 15, 1–10 (1972a)

    Google Scholar 

  • Bron, S., Venema, G.: Ultraviolet inactivation and excision-repair in Bacillus subtilis. III. Sensitized photoinactivation of transforming DNA, and the effect of thymine dimers on differential marker inactivation and differential marker repair. Mutation Res. 15, 377–393 (1972b)

    Google Scholar 

  • Dussoix, D., Arber, W.: Host specificity of DNA produced by Escherichia coli. IV. Host specificity of infectious DNA from bacteriophage lambda. J. molec. Biol. 11, 238–246 (1965)

    Google Scholar 

  • Garfin, D.E., Goodman, H.M.: Nucleotide sequences at the cleavage sites of two restriction endonucleases from Haemophilus parainfluenzae. Biochem. biophys. Res. Comm. 59, 108–116 (1974)

    Google Scholar 

  • Goodgal, S.H., Gromkova, R.: On the role of restriction enzymes of Haemophilus influenzae in transformation and transfection. In: Abstracts 2nd Eur. Meeting Transformation and Transfection, Polish Acad. Sci., Cracow. 49–52 (1974)

  • Gromkova, R., Goodgal, S.H.: Action of Haemophilus endodeoxyribonuclease on biologically active deoxyribonucleic acid. J. Bact. 109, 987–992 (1972)

    Google Scholar 

  • Hedgpeth, J., Goodman, H.M., Boyer, H.W.: DNA nucleotide sequence restricted by the RI endonuclease. Proc. nat. Acad. Sci. (Wash.) 69, 3448–3452 (1972)

    Google Scholar 

  • Hirokawa, H.: Transfecting deoxyribonucleic acid of Bacillus bacteriophage ϕ29 that is protease sensitive. Proc. nat. Acad. Sci. (Wash.) 69, 1555–1559 (1972)

    Google Scholar 

  • Jackson, D.A., Symons, R.H., Berg, P.: Biochemical method for inserting new genetic information into DNA of Simian Virus 40: Circular SV40 DNA molecules containing lambda phage genes and the galactose operon of Escherichia coli. Proc. nat. Acad. Sci. (Wash.) 69, 2904–2909 (1972)

    Google Scholar 

  • Kelly, T.J., Smith, H.O.: A restriction enzyme from Haemophilus influenzae. II. Base sequence of the recognition site. J. molec. Biol. 51, 393–409 (1970)

    Google Scholar 

  • Lai, J., Nathaus, D.: Mapping temperature-sensitive mutants of simian-virus 40: Rescue of mutants by fragments of viral DNA. Virology 60, 466–475 (1974)

    Google Scholar 

  • Linn, S., Arber, W.: Host specificity of DNA produced by Escherichia coli. X. In vitro restriction of phage fd replicative form. Proc. nat. Acad. Sci. (Wash.) 59, 1300–1306 (1968)

    Google Scholar 

  • Lowry, O.H., Rosebrough, N.J., Farr, A.L., Randall, R.J.: Protein measurement with the folin phenol reagent. J. biol. Chem. 193, 265–275 (1951)

    Google Scholar 

  • Marx, J.L.: Restriction enzymes: New Tools for studying DNA. Science 180, 482–485 (1973)

    Google Scholar 

  • Meselson, M., Yuan, R.: DNA restriction enzyme from E. coli. Nature (Lond.) 217, 1110–1114 (1968)

    Google Scholar 

  • Meselson, M., Yuan, R., Heywood, J.: Restriction and modification of DNA. Ann. Rev. Biochem. 41, 447–466 (1972)

    Google Scholar 

  • Middleton, J.H., Edgell, M.H., Hutchinson, C.A. III: Specific fragments of ϕX174 deoxyribonucleic acid produced by a restriction enzyme from Haemophilus aegyptius, endonuclease Z. J. Virol. 10, 42–50 (1972)

    Google Scholar 

  • Morrow, J.F., Cohen, S.N., Chang, A.C.Y., Boyer, H.W., Goodman, H.M., Helling, R.B.: Proc. nat. Acad. Sci. (Wash.) 71, 1743–1747 (1972)

    Google Scholar 

  • Murray, K.: Nucleotide sequence analysis with polynucleotide kinase and nucleotide “Mapping” methods. Biochem. J. 131, 569–583 (1973)

    Google Scholar 

  • Murray, N.E., Murray, K.: Manipulation of restriction targets and DNA fragments of phage λ: Recpetor chromosomes for fragments of DNA. Nature (Lond.) 251, 476–481 (1974).

    Google Scholar 

  • Murray, K., Old, R.W.: The primary structure of DNA. Proc. Nucleic Acid Res. Mol. Biol. 14, 117–185, Academic Press Inc. (1974)

    Google Scholar 

  • Old, R.W., Murray, K., Roizes, G.: Recognition sequence of restriction endonuclease III from Haemophilus influenzae. J. molec. Biol. 92, 331–339 (1975)

    Google Scholar 

  • Okubo, S., Romig, W.R.: Imparied transformability of Bacillus subtilis mutant sentitive to mitomycin C and ultraviolet radiation. J. molec. Biol. 15, 440–454 (1966)

    Google Scholar 

  • Rambach, A., Tiollais, P.: Bacteriophage λ having Eco R1 endonuclease sites only in the non-essential region of the genome. Proc. nat. Acad. Sci. (Wash.) 71, 3927–3930 (1974)

    Google Scholar 

  • Rottländer, E., Trautner, T.A.: Genetic and transfection studies with B. subtilis phage SP50. I. Phage mutants with restricted growth on B. subtilis strain 168. Molec. gen. Genet. 108, 47–60 (1970)

    Google Scholar 

  • Sharp, P.A., Sugden, B., Sambrook, J.: Detection of two restriction endonuclease activities in Haemophilus parainfluenzae using analytical agarose-ethidium bromide electrophoresis. Biochemistry (Wash.) 12, 3055–3063 (1973)

    Google Scholar 

  • Smith, H.O., Nathans, D.: A suggested nomenclature for bacterial host modification and restriction systems and their enzymes. J. molec. Biol. 81, 419–423 (1973)

    Google Scholar 

  • Smith, H.O., Wilcox, K.W.: A restriction enzyme from Haemophilus influenzae. I. Purification and general properties. J. molec. Biol. 51, 379–391 (1970)

    Google Scholar 

  • Sugisaki, H., Takanami, M.: DNA sequence restricted by restriction endonuclease AP from Haemophilus aphirophilus. Nature (Lond.) New Biol. 246, 138–140 (1973)

    Google Scholar 

  • Takano, T., Watanabe, T., Fukasawa, T.: Mechanism of host-controlled restriction of bacteriophage λ by R factors in Escherichia coli K12. Virology 34, 290–302 (1968)

    Google Scholar 

  • Thomas, M., Cameron, J.R., Davis, R.W.: Viable molecular hybrids of bacteriophage λ and eukaryotic DNA. Proc. nat. Acad. Sci. (Wash.) 71, 4579–4583 (1974)

    Google Scholar 

  • Trautner, T.A., Pawlek, B., Bron, S., Anagnotopoulos, C.: Restriction and modification in B. subtilis. Biological Aspects. Molec. gen. Genet. 109, 181–191 (1974)

    Google Scholar 

  • Wilson, G.A., Young, F.E.: Intergenotic transformation of the Bacillus subtilis genospecies. J. Bact. 111, 705–716 (1972)

    Google Scholar 

  • Wilson, G.A., Young, F.E.: Restriction of homologous and heterologous transfection and infection in Bacillus subtilis. In: Abstracts 2nd Eur. Meeting Transformation and Transfection, Polish Acad. Sci. Cracow, 141 (1974)

  • Yamamoto, K.R., Alberts, B.M., Benzinger, R., Lawborne, L., Treiber, G.: Rapid bacteriophage sedimentation in the presence of polyethylene glycol and its application to large-scale virus purification. Virology 40, 734–744 (1970)

    Google Scholar 

  • Yoshimori, R.N.: A genetical and biochemical analysis of the restriction and modification of DNA by resistance transfer factors. 73 pp. Ph.D. thesis, University of California (1971)

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Authors and Affiliations

  1. Department of Molecular Biology, University of Edinburgh, Edinburgh, Scotland

    Sierd Bron

  2. Department of Genetics, Biological Centre, Haren (Gn), The Netherlands

    Sierd Bron

  3. Department of Molecular Biology, University of Edinburgh, EH9 3JR, Edinburgh, Scotland

    Kenneth Murray

  4. Max-Planck-Institut für Molekulare Genetik, Ihnestr. 63-73, D-1000, Berlin 33, Germany

    Thomas A. Trautner

Authors
  1. Sierd Bron
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  2. Kenneth Murray
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  3. Thomas A. Trautner
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Communcated by W. Arber

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Bron, S., Murray, K. & Trautner, T.A. Restriction and modification in B. subtilis . Molec. Gen. Genet. 143, 13–23 (1975). https://doi.org/10.1007/BF00269416

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  • DOI: https://doi.org/10.1007/BF00269416

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