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. 1985 Oct 25;13(20):7171–7182. doi: 10.1093/nar/13.20.7171

Purification of Mbo II methylase (GAAGmA) from Moraxella bovis: site specific cleavage of DNA at nine and ten base pair sequences.

M McClelland, M Nelson, C R Cantor
PMCID: PMC322036  PMID: 2997742

Abstract

The restriction modification methylase M. Mbo II has been purified using a sensitive oligonucleotide linker assay. The enzyme methylates the Mbo II recognition sequence* GAAGA at adenine to produce GAAGmA. M. Mbo II can be used in conjunction with the methylation dependent restriction endonuclease Dpn I (GmATC) to produce cleavage at the 10 base sequence GAAGATCTTC. When M. Mbo II is used in combination with M. Cla I (ATCGATCGAT), cleavage by Dpn I occurs at the four ten base sequences GAAGATCTTC, GAAGATCGAT, ATCGATCTTC and ATCGATCGAT, which is equivalent to a nine base recognition site. The use of combinations of adenine methylases and Dpn I to generate highly selective DNA cleavages at a variety of sequences up to fourteen base pairs is discussed.

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Selected References

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  1. Bibb M. J., Van Etten R. A., Wright C. T., Walberg M. W., Clayton D. A. Sequence and gene organization of mouse mitochondrial DNA. Cell. 1981 Oct;26(2 Pt 2):167–180. doi: 10.1016/0092-8674(81)90300-7. [DOI] [PubMed] [Google Scholar]
  2. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brooks J. E., Roberts R. J. Modification profiles of bacterial genomes. Nucleic Acids Res. 1982 Feb 11;10(3):913–934. doi: 10.1093/nar/10.3.913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brown N. L., Hutchison C. A., 3rd, Smith M. The specific non-symmetrical sequence recognized by restriction endonuclease MboII. J Mol Biol. 1980 Jun 15;140(1):143–148. doi: 10.1016/0022-2836(80)90360-5. [DOI] [PubMed] [Google Scholar]
  5. Bächi B., Reiser J., Pirrotta V. Methylation and cleavage sequences of the EcoP1 restriction-modification enzyme. J Mol Biol. 1979 Feb 25;128(2):143–163. doi: 10.1016/0022-2836(79)90123-2. [DOI] [PubMed] [Google Scholar]
  6. Gelinas R. E., Myers P. A., Roberts R. J. Two sequence-specific endonucleases from Moraxella bovis. J Mol Biol. 1977 Jul;114(1):169–179. doi: 10.1016/0022-2836(77)90290-x. [DOI] [PubMed] [Google Scholar]
  7. Hosbach H. A., Silberklang M., McCarthy B. J. Evolution of a D. melanogaster glutamate tRNA gene cluster. Cell. 1980 Aug;21(1):169–178. doi: 10.1016/0092-8674(80)90124-5. [DOI] [PubMed] [Google Scholar]
  8. Joyce C. M., Kelley W. S., Grindley N. D. Nucleotide sequence of the Escherichia coli polA gene and primary structure of DNA polymerase I. J Biol Chem. 1982 Feb 25;257(4):1958–1964. [PubMed] [Google Scholar]
  9. McClelland M., Kessler L. G., Bittner M. Site-specific cleavage of DNA at 8- and 10-base-pair sequences. Proc Natl Acad Sci U S A. 1984 Feb;81(4):983–987. doi: 10.1073/pnas.81.4.983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. McClelland M., Nelson M. The effect of site specific methylation on restriction endonuclease digestion. Nucleic Acids Res. 1985;13 (Suppl):r201–r207. doi: 10.1093/nar/13.suppl.r201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. McClelland M. Purification and characterization of two new modification methylases: MClaI from Caryophanon latum L and MTaqI from Thermus aquaticus YTI. Nucleic Acids Res. 1981 Dec 21;9(24):6795–6804. doi: 10.1093/nar/9.24.6795. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Nelson M., Christ C., Schildkraut I. Alteration of apparent restriction endonuclease recognition specificities by DNA methylases. Nucleic Acids Res. 1984 Jul 11;12(13):5165–5173. doi: 10.1093/nar/12.13.5165. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Roberts R. J. Restriction and modification enzymes and their recognition sequences. Nucleic Acids Res. 1985;13 (Suppl):r165–r200. doi: 10.1093/nar/13.suppl.r165. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Sato S., Nakazawa K., Shinomiya T. A DNA methylase from Thermus thermophilus HB8. J Biochem. 1980 Sep;88(3):737–747. doi: 10.1093/oxfordjournals.jbchem.a133026. [DOI] [PubMed] [Google Scholar]
  15. Schwartz D. C., Cantor C. R. Separation of yeast chromosome-sized DNAs by pulsed field gradient gel electrophoresis. Cell. 1984 May;37(1):67–75. doi: 10.1016/0092-8674(84)90301-5. [DOI] [PubMed] [Google Scholar]
  16. van Ormondt H., Lautenberger J. A., Linn S., de Waard A. Methylated oligonucleotides derived from bacteriophage fd RF-DNA modified in vitro by E. coli B modification methylase. FEBS Lett. 1973 Jul 1;33(2):177–180. doi: 10.1016/0014-5793(73)80186-3. [DOI] [PubMed] [Google Scholar]

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