Rapid and Efficient Plasmid Construction by Homologous Recombination in Yeast.

Détails

ID Serval
serval:BIB_B722E812F733
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Rapid and Efficient Plasmid Construction by Homologous Recombination in Yeast.
Périodique
Cold Spring Harbor protocols
Auteur⸱e⸱s
van Leeuwen J., Andrews B., Boone C., Tan G.
ISSN
1559-6095 (Electronic)
ISSN-L
1559-6095
Statut éditorial
Publié
Date de publication
01/09/2015
Volume
2015
Numéro
9
Pages
pdb.prot085100
Langue
anglais
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: epublish
Résumé
The cloning of DNA fragments is a fundamental aspect of molecular biology. Traditional DNA cloning techniques rely on the ligation of an insert and a linearized plasmid that have been digested with restriction enzymes and the subsequent introduction of the ligated DNA into Escherichia coli for propagation. However, this method is limited by the availability of restriction sites, which often becomes problematic when cloning multiple or large DNA fragments. Furthermore, using traditional methods to clone multiple DNA fragments requires experience and multiple laborious steps. In this protocol, we describe a simple and efficient cloning method that relies on homologous recombination in the yeast Saccharomyces cerevisiae to assemble multiple DNA fragments, with 30-bp homology regions between the fragments, into one sophisticated construct. This method can easily be extended to clone plasmids for other organisms, such as bacteria, plants, and mammalian cells.
Mots-clé
Cloning, Molecular/methods, Homologous Recombination, Molecular Biology/methods, Plasmids/isolation & purification, Saccharomyces cerevisiae/genetics
Pubmed
Open Access
Oui
Création de la notice
22/01/2019 16:04
Dernière modification de la notice
21/08/2019 5:35
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