A two-plasmid bacterial selection system for characterization and engineering of homing endonucleases

Ning Sun, Huimin Zhao

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Homing endonucleases recognize long DNA sequences and generate site-specific DNA double-stranded breaks. They can serve as a powerful genomic modification tool in various industrial and biomedical applications. Here, we describe a two-plasmid bacterial selection system for characterization and engineering of homing endonucleases. This selection system couples the DNA cleavage activity of a homing endonuclease with the survival of host cells. Therefore, it can be used for assaying in vivo activity of homing endonucleases. Moreover, due to its high sensitivity, it can be applied for directed evolution of homing endonucleases with altered sequence specificity.

Original languageEnglish (US)
Title of host publicationHoming Endonucleases
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Pages87-96
Number of pages10
ISBN (Print)9781627039673
DOIs
StatePublished - 2014

Publication series

NameMethods in Molecular Biology
Volume1123
ISSN (Print)1064-3745

Fingerprint

Endonucleases
Plasmids
DNA Cleavage
Double-Stranded DNA Breaks
Cell Survival

Keywords

  • Directed evolution
  • Gene targeting
  • Gene therapy
  • Homing endonucleases
  • Protein engineering

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Sun, N., & Zhao, H. (2014). A two-plasmid bacterial selection system for characterization and engineering of homing endonucleases. In Homing Endonucleases: Methods and Protocols (pp. 87-96). (Methods in Molecular Biology; Vol. 1123). Humana Press Inc.. https://doi.org/10.1007/978-1-62703-968-0_7

A two-plasmid bacterial selection system for characterization and engineering of homing endonucleases. / Sun, Ning; Zhao, Huimin.

Homing Endonucleases: Methods and Protocols. Humana Press Inc., 2014. p. 87-96 (Methods in Molecular Biology; Vol. 1123).

Research output: Chapter in Book/Report/Conference proceedingChapter

Sun, N & Zhao, H 2014, A two-plasmid bacterial selection system for characterization and engineering of homing endonucleases. in Homing Endonucleases: Methods and Protocols. Methods in Molecular Biology, vol. 1123, Humana Press Inc., pp. 87-96. https://doi.org/10.1007/978-1-62703-968-0_7
Sun N, Zhao H. A two-plasmid bacterial selection system for characterization and engineering of homing endonucleases. In Homing Endonucleases: Methods and Protocols. Humana Press Inc. 2014. p. 87-96. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-62703-968-0_7
Sun, Ning ; Zhao, Huimin. / A two-plasmid bacterial selection system for characterization and engineering of homing endonucleases. Homing Endonucleases: Methods and Protocols. Humana Press Inc., 2014. pp. 87-96 (Methods in Molecular Biology).
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