Molecular and General Genetics MGG

, Volume 260, Issue 2, pp 226–231

Cloning and expression of the Apa LI, Nsp I, Nsp HI, Sac I, Sca I, and Sap I restriction-modification systems in Escherichia coli

  • S.-y. Xu
  • J.-p. Xiao
  • L. Ettwiller
  • M. Holden
  • J. Aliotta
  • C. L. Poh
  • M. Dalton
  • D. P. Robinson
  • T. R. Petronzio
  • L. Moran
  • M. Ganatra
  • J. Ware
  • B. Slatko
  • J. Benner
ORIGINAL PAPER

DOI: 10.1007/s004380050890

Cite this article as:
Xu, S., Xiao, J., Ettwiller, L. et al. Mol Gen Genet (1998) 260: 226. doi:10.1007/s004380050890

Abstract

The genes encoding the ApaLI (5′-G^TGCAC-3′), NspI (5′-RCATG^Y-3′), NspHI (5′-RCATG^Y-3′), SacI (5′-GAGCT^C-3′), SapI (5′-GCTCTTCN1^-3′, 5′-^N4GAAGAGC-3′) and ScaI (5′-AGT^ACT-3′) restriction-modification systems have been cloned in E.␣coli. Amino acid sequence comparison of M.ApaLI, M.NspI, M.NspHI, and M.SacI with known methylases indicated that they contain the ten conserved motifs characteristic of C5 cytosine methylases. NspI and NspHI restriction-modification systems are highly homologous in amino acid sequence. The C-termini of the NspI and NlaIII (5′-CATG-3′) restriction endonucleases share significant similarity. 5mC modification of the internal C in a SacI site renders it resistant to SacI digestion. External 5mC modification of a SacI site has no effect on SacI digestion. N4mC modification of the second base in the sequence 5′-GCTCTTC-3′ blocks SapI digestion. N4mC modification of the other cytosines in the SapI site does not affect SapI digestion. N4mC modification of ScaI site blocks ScaI digetion. A DNA invertase homolog was found adjacent to the ApaLI restriction-modification system. A DNA transposase subunit homolog was found upstream of the SapI restriction endonuclease gene.

Key words Restriction endonuclease Methylase selection Gene expression DNA methylation Recombinant DNA 

Copyright information

© Springer-Verlag Berlin Heidelberg 1998

Authors and Affiliations

  • S.-y. Xu
    • 1
  • J.-p. Xiao
    • 1
  • L. Ettwiller
    • 1
  • M. Holden
    • 2
  • J. Aliotta
    • 3
  • C. L. Poh
    • 4
  • M. Dalton
    • 1
  • D. P. Robinson
    • 1
  • T. R. Petronzio
    • 1
  • L. Moran
    • 1
  • M. Ganatra
    • 1
  • J. Ware
    • 1
  • B. Slatko
    • 1
  • J. Benner
    • 1
  1. 1.New England Biolabs, Inc., 32 Tozer Road, Beverly MA 01915, USA e-mail: xus@neb.com Tel.: +1-978-927-5054; Fax: +1-978-921-1350 GB
  2. 2.Brown University, Box 1258, Providence RI 02912, USAUS
  3. 3.Tufts University Medical School 136 Harrison Ave. Boston, MA 02111, USAUS
  4. 4.Dept. of Microbiology, Faculty of Medicine National University of Singapore Lower Kent Ridge Rd. Singapore 0511SG

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