A new method for constructing linker scanning mutants | Zendy

Bruno Luckow | Zendy; Rainer Renkawitz | Zendy; Günther Schütz | Zendy

AI Assistant Blog Pricing

Home ZAIA Blog

Open Access

A new method for constructing linker scanning mutants

Author(s) -

Bruno Luckow,

Rainer Renkawitz,

Günther Schütz

Publication year - 1987

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/15.2.417

Subject(s) - linker , biology , plasmid , exonuclease iii , mutant , exonuclease , linker dna , nuclease , dna , ap site , cleavage (geology) , microbiology and biotechnology , genetics , escherichia coli , dna polymerase , gene , endonuclease , chromatin , nucleosome , computer science , operating system , paleontology , fracture (geology)

A new procedure for the construction of linker scanning mutants is described. A plasmid containing the target DNA is randomly linearized and slightly shortened by a novel combination of established methods. After partial apurination with formic acid a specific nick or small gap is introduced at the apurinic site by exonuclease III, followed by nuclease S1 cleavage of the strand opposite the nick/gap. Synthetic linkers are ligated to the ends and plasmids having the linker inserted in the target DNA are enriched. Putative linker scanning mutants are identified by their topoisomer patterns after relaxation with topoisomerase I. This technique allows the distinction of plasmids differing in length by a single basepair. We have used this rapid and efficient strategy to generate a set of 32 linker scanning mutants covering the chicken lysozyme promoter from -208 to +15.

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