Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1979 Mar;137(3):1299–1307. doi: 10.1128/jb.137.3.1299-1307.1979

NgoII, a restriction endonuclease from Neisseria gonorrhoeae.

D J Clanton, W S Riggsby, R V Miller
PMCID: PMC218313  PMID: 35516

Abstract

EndoR . NgoII, a class II restriction endonuclease isolated from Neisseria gonorrhoeae, was purified to electrophoretic homogeneity. We were able to separate it from another restriction endonuclease of N. gonorrhoeae, NgoI, by phosphocellulose chromatography. NgoII is an isoschizomer of HaeIII, a restriction endonuclease of Haemophilus aegyptius, and was found to recognize the deoxyribonucleic acid nucleotide base sequence GGCC. NgoII was able to digest phage lambda deoxyribonucleic acid over a wide pH range, with optimal activity at pH 8.5. The enzyme has an absolute requirement for Mg2+; maximal enzyme activity was observed at 1 mM Mg2+. The active enzyme has a molecular weight of 65,000 and appears to be composed of six subunits of identical molecular weight (11,000). No methylase activity could be detected in the purified enzyme preparation.

Full text

PDF
1299

Images in this article

1302Image
on p.1302
1303Image
on p.1303
1303Image
on p.1303

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Andrews P. Estimation of the molecular weights of proteins by Sephadex gel-filtration. Biochem J. 1964 May;91(2):222–233. doi: 10.1042/bj0910222. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Arwert F., Rutberg L. Restriction and modification in Bacillus subtilis. Induction of a modifying activity in Bacillus subtilis 168. Mol Gen Genet. 1974;133(2):175–177. doi: 10.1007/BF00264838. [DOI] [PubMed] [Google Scholar]
  3. Boyer H. W. Restriction and modification of DNA: enzymes and substrates. Introductory remarks. Fed Proc. 1974 May;33(5):1125–1127. [PubMed] [Google Scholar]
  4. Catterall J. F., Welker N. E. Isolation and properties of a thermostable restriction endonuclease (ENDO R-Bst1503). J Bacteriol. 1977 Feb;129(2):1110–1120. doi: 10.1128/jb.129.2.1110-1120.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Clanton D. J., Woodward J. M., Miller R. V. Identification of a new sequence-specific endonuclease, NgoII, from Neisseria gonorrhoeae. J Bacteriol. 1978 Jul;135(1):270–273. doi: 10.1128/jb.135.1.270-273.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Eisenstein B. I., Sox T., Biswas G., Blackman E., Sparling P. F. Conjugal transfer of the gonococcal penicillinase plasmid. Science. 1977 Mar 11;195(4282):998–1000. doi: 10.1126/science.402693. [DOI] [PubMed] [Google Scholar]
  7. Eskin B., Linn S. The deoxyribonucleic acid modification and restriction enzymes of Escherichia coli B. II. Purification, subunit structure, and catalytic properties of the restriction endonuclease. J Biol Chem. 1972 Oct 10;247(19):6183–6191. [PubMed] [Google Scholar]
  8. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  9. Lautenberger J. A., Linn S. The deoxyribonucleic acid modification and restriction enzymes of Escherichia coli B. I. Purification, subunit structure, and catalytic properties of the modification methylase. J Biol Chem. 1972 Oct 10;247(19):6176–6182. [PubMed] [Google Scholar]
  10. Linn S., Lautenberger J. A., Eskin B., Lackey D. Host-controlled restriction and modification enzymes of Escherichia coli B. Fed Proc. 1974 May;33(5):1128–1134. [PubMed] [Google Scholar]
  11. Miller R. V., Clark A. J. Purification and properties of two deoxyribonucleases of Pseudomonas aeruginosa. J Bacteriol. 1976 Aug;127(2):794–802. doi: 10.1128/jb.127.2.794-802.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Murray N. E., Batten P. L., Murray K. Restriction of bacteriophage lambda by Escherichia coli K. J Mol Biol. 1973 Dec 15;81(3):395–407. doi: 10.1016/0022-2836(73)90149-6. [DOI] [PubMed] [Google Scholar]
  13. Reisner A. H., Nemes P., Bucholtz C. The use of Coomassie Brilliant Blue G250 perchloric acid solution for staining in electrophoresis and isoelectric focusing on polyacrylamide gels. Anal Biochem. 1975 Apr;64(2):509–516. doi: 10.1016/0003-2697(75)90461-3. [DOI] [PubMed] [Google Scholar]
  14. Roberts M., Elwell L. P., Falkow S. Molecular characterization of two beta-lactamase-specifying plasmids isolated from Neisseria gonorrhoeae. J Bacteriol. 1977 Aug;131(2):557–563. doi: 10.1128/jb.131.2.557-563.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Roberts M., Falkow S. Plasmid-mediated chromosomal gene transfer in Neisseria gonorrhoeae. J Bacteriol. 1978 Apr;134(1):66–70. doi: 10.1128/jb.134.1.66-70.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Roberts R. J. Restriction endonucleases. CRC Crit Rev Biochem. 1976 Nov;4(2):123–164. doi: 10.3109/10409237609105456. [DOI] [PubMed] [Google Scholar]
  17. Sharp P. A., Sugden B., Sambrook J. Detection of two restriction endonuclease activities in Haemophilus parainfluenzae using analytical agarose--ethidium bromide electrophoresis. Biochemistry. 1973 Jul 31;12(16):3055–3063. doi: 10.1021/bi00740a018. [DOI] [PubMed] [Google Scholar]
  18. Sparling P. F. Genetic transformation of Neisseria gonorrhoeae to streptomycin resistance. J Bacteriol. 1966 Nov;92(5):1364–1371. doi: 10.1128/jb.92.5.1364-1371.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Stiffler P. W., Lerner S. A., Bohnhoff M., Morello J. A. Plasmid deoxyribonucleic acid in clinical isolates of Neisseria gonorrhoeae. J Bacteriol. 1975 Jun;122(3):1293–1300. doi: 10.1128/jb.122.3.1293-1300.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES