An in vivo selection system for homing endonuclease activity | Zendy

Michael Gruen | Zendy

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An in vivo selection system for homing endonuclease activity

Author(s) -

Michael Gruen

Publication year - 2002

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/30.7.e29

Subject(s) - homing endonuclease , biology , endonuclease , homing (biology) , dna , cleavage (geology) , genetics , restriction enzyme , mutant , computational biology , microbiology and biotechnology , gene , ecology , paleontology , fracture (geology)

Homing endonucleases are enzymes that catalyze the highly sequence-specific cleavage of DNA. We have developed an in vivo selection in Escherichia coli that links cell survival with homing endonuclease-mediated DNA cleavage activity and sequence specificity. Using this selection, wild-type and mutant variants of three homing endonucleases were characterized without requiring protein purification and in vitro analysis. This selection system may facilitate the study of sequence-specific DNA cleaving enzymes, and selections based on this work may enable the evolution of homing endonucleases with novel activities or specificities.

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