Vectors with hidden cloning sites

Détails

ID Serval
serval:BIB_9CBE40D674E5
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Vectors with hidden cloning sites
Périodique
Biochemical and Biophysical Research Communications
Auteur⸱e⸱s
Welker  E., Varadi  A.
ISSN
0006-291X (Print)
Statut éditorial
Publié
Date de publication
05/2000
Volume
271
Numéro
2
Pages
534-6
Notes
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: May 10
Résumé
A general strategy is described for using the cleavage site of restriction enzymes in vectors for cloning regardless of how many sites the given enzymes have in the vector. The application of this method allows one to open any vector at its cloning site with protruding ends which can be compatible with almost every commercially available Class II restriction enzyme. By employing this method, the laborious construction of new vectors can be simplified considerably. This general strategy is based on the known ability of Class IIS restriction enzymes to cut any sequence located outside of their recognition site; the introduction of a linker containing recognition site(s) for Class IIS restriction enzyme(s), not present originally in the vector, gives rise to the possibility of opening the vector so as to produce overhangs of arbitrary sequence. In particular, when a symmetrical short sequence representing the protruding end of any Class II enzyme is situated at the cutting position of the Class IIS enzyme, cleavage with the Class IIS enzyme exposes the hitherto hidden, "unique" cloning site. This technique is demonstrated by cloning the cDNA of the multidrug resistance protein to an expression vector.
Mots-clé
Animals Cloning, Molecular/*methods DNA, Complementary/metabolism Deoxyribonucleases, Type II Site-Specific/metabolism *Genetic Vectors Humans Insects
Pubmed
Web of science
Création de la notice
24/01/2008 15:40
Dernière modification de la notice
20/08/2019 16:03
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