A Novel System for the Rapid Generation of Precise DNA Deletions | Zendy

Ian McCaffery | Zendy; Brian D. Williamson | Zendy; C. L. Rutherford | Zendy

AI Assistant Blog Pricing

Home ZAIA Blog

Open Access

A Novel System for the Rapid Generation of Precise DNA Deletions

Author(s) -

Ian McCaffery,

Brian D. Williamson,

C. L. Rutherford

Publication year - 1996

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/24.24.5048

Subject(s) - biology , restriction enzyme , dna , recognition sequence , plasmid , ligation , genetics , dna ligase , microbiology and biotechnology , restriction map , sequencing by ligation , restriction site , cleavage (geology) , restriction fragment , in vitro recombination , gene , molecular cloning , peptide sequence , genomic library , base sequence , paleontology , fracture (geology)

To generate DNA deletions, a tandem array of class IIS restriction enzyme recognition sites was cloned into a plasmid. The recognition sites were arranged so that each enzyme cleaves at a different site within an adjacent target sequence. Digestion with both enzymes followed by end repair and ligation resulted in the deletion of DNA between the two sites of cleavage. Because both recognition sites are preserved following deletion, it was found that sequential deletions could be generated using cycles of restriction enzyme digestion, end repair and ligation. Therefore, this system represents a valuable tool in the definition of functional DNA sequences.

The content you want is available to Zendy users.

Already have an account? Click here to sign in.

Having issues? You can contact us here

Accelerating Research