Length of flanking homologous regions required for gene targeting through homologous recombination in yeast

Value >= bp Range: ≥30 bp
Organism Budding yeast Saccharomyces cerevisiae
Reference Manivasakam P, Weber SC, McElver J, Schiestl RH. Micro-homology mediated PCR targeting in Saccharomyces cerevisiae. Nucleic Acids Res. 1995, 23:2799-800. p.2800 right column bottom paragraphPubMed ID7651842
Method P.2799 left column top paragraph: "In the present study, [investigators] determined the amount of homology required for targeted integration of DNA fragments into the yeast genome. The procedure described here facilitates the manipulation of the yeast genome and eliminates the need to clone sequences homologous to a target site. In addition, this method is useful for applications in which only limited sequence information of the target is available. The procedure comprises of: (i) production of PCR primers to amplify a selectable marker containing flanking homology to the target of choice (ii) transformation of yeast cells and (iii) selection of integrants."
Comments P.2800 right column bottom paragraph: "In summary, [investigators] have defined the minimum amount of homology required for efficient homologous integration in S. cerevisiae. Homology of 30 bp on each side of a selectable marker is sufficient to obtain a large fraction of targeted integration events. This information can be applied to the economical design of primers for yeast genome modification by microhomology mediated PCR targeting."
Entered by Irina
ID 101986