Chimeric restriction enzyme: Gal4 fusion to Fokl cleavage domain

Yang Gyun Kim, Jeff Smith, Srinivasan Chandrasegaran, Srinivasan Chandrasegaran

Research output: Contribution to journalArticle

Abstract

Gal4, a yeast protein, activates transcription of genes required for metabolism of galactose and melibiose. It binds as a dimer to a consensus palindromic 17-base pair DNA sequence. It is a member of the third family of proteins that contain zinc-mediated peptide loops that interact specifically with nucleic acids. Gal4 has a very distinctive zinc coordination profile and mode of DNA-binding. Here, we report the creation of a novel site-specific endonuclease by linking the N-terminal 147 amino acids of Gal4 to the cleavage domain of Fokl endonuclease. The fusion protein is active and under optimal conditions, binds to a 17 bp consensus DNA site and cleaves near this site. As expected, the cleavage occurs on either side of the consensus binding site(s).

Original languageEnglish (US)
Pages (from-to)489-495
Number of pages7
JournalBiological Chemistry
Volume379
Issue number4-5
StatePublished - Apr 1998
Externally publishedYes

Fingerprint

Endonucleases
Zinc
Fusion reactions
Melibiose
Fungal Proteins
DNA sequences
DNA
Enzymes
Transcription
Galactose
Metabolism
Base Pairing
Dimers
Nucleic Acids
Consensus
Proteins
Genes
Binding Sites
Amino Acids
Peptides

Keywords

  • Chimeric restriction endonuclease
  • Flavobacterium okeanokoites
  • Hybrid restriction enzymes
  • Leucine zipper motifs
  • Recognition and cleavage domains
  • Zinc binding proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Kim, Y. G., Smith, J., Chandrasegaran, S., & Chandrasegaran, S. (1998). Chimeric restriction enzyme: Gal4 fusion to Fokl cleavage domain. Biological Chemistry, 379(4-5), 489-495.

Chimeric restriction enzyme : Gal4 fusion to Fokl cleavage domain. / Kim, Yang Gyun; Smith, Jeff; Chandrasegaran, Srinivasan; Chandrasegaran, Srinivasan.

In: Biological Chemistry, Vol. 379, No. 4-5, 04.1998, p. 489-495.

Research output: Contribution to journalArticle

Kim, YG, Smith, J, Chandrasegaran, S & Chandrasegaran, S 1998, 'Chimeric restriction enzyme: Gal4 fusion to Fokl cleavage domain', Biological Chemistry, vol. 379, no. 4-5, pp. 489-495.
Kim, Yang Gyun ; Smith, Jeff ; Chandrasegaran, Srinivasan ; Chandrasegaran, Srinivasan. / Chimeric restriction enzyme : Gal4 fusion to Fokl cleavage domain. In: Biological Chemistry. 1998 ; Vol. 379, No. 4-5. pp. 489-495.
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