Patent ID: 6090554
Filing Date: 2000-07-18
Classification: A01K,C12N

Abstract:
A method for preparing gene targeting vectors in yeast, the method comprising:a) preparing a shuttle vector comprising a first yeast selectable marker, a bacterial selectable marker and a fragment of genomic DNA containing at least part of a gene to be targeted;b) preparing a specific engineered fragment (SEF) comprising a marker cassette, the marker cassette comprising a second yeast selectable marker different from the first yeast selectable marker, a selectable marker capable of expression in mammalian embryonic stem cells said marker cassette being flanked on each side by mammalian gene-specific flanking sequences homologous to a portion of the gene to be targeted;c) transforming yeast cells with the shuttle vector of step a) and with the SEF of step b), and allowing said shuttle vector and said SEF to recombine by homologous recombination;d) selecting the transformed yeast cells for expression of said first and second yeast selectable markers; ande) isolating the targeting vector produced by recombination between the shuttle vector and the SEF from the yeast cells selected in step d).