The AplI restriction-modification system in an edible cyanobacterium, Arthrospira (Spirulina) platensis NIES-39, recognizes the nucleotide sequence 5'-CTGCAG-3'

Biosci Biotechnol Biochem. 2013;77(4):782-8. doi: 10.1271/bbb.120919. Epub 2013 Apr 7.

Abstract

The degradation of foreign DNAs by restriction enzymes in an edible cyanobacterium, Arthrospira platensis, is a potential barrier for gene-transfer experiments in this economically valuable organism. We overproduced in Escherichia coli the proteins involved in a putative restriction-modification system of A. platensis NIES-39. The protein produced from the putative type II restriction enzyme gene NIES39_K04640 exhibited an endonuclease activity that cleaved DNA within the sequence 5'-CTGCAG-3' between the A at the fifth position and the G at the sixth position. We designated this enzyme AplI. The protein from the adjacent gene NIES39_K04650, which encodes a putative DNA (cytosine-5-)-methyltransferase, rendered DNA molecules resistant to AplI by modifying the C at the fourth position (but not the C at the first position) in the recognition sequence. This modification enzyme, M.AplI, should be useful for converting DNA molecules into AplI-resistant forms for use in gene-transfer experiments. A summary of restriction enzymes in various Arthrospira strains is also presented in this paper.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Binding Sites
  • DNA Restriction Enzymes / chemistry
  • DNA Restriction Enzymes / isolation & purification
  • DNA Restriction Enzymes / metabolism*
  • DNA, Bacterial / genetics*
  • DNA, Bacterial / metabolism*
  • Escherichia coli / genetics
  • Food Microbiology*
  • Protein Binding
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism*
  • Spirulina / enzymology*
  • Spirulina / genetics
  • Substrate Specificity

Substances

  • DNA, Bacterial
  • Recombinant Proteins
  • DNA Restriction Enzymes