<i>Oncotarget</i> V9N20 (IMAGE)

Impact Journals LLC

<i>Oncotarget</i> V9N20

Caption

The diagram showing a general scheme of TAR cloning of a region of interest (in red) from total genomic DNA (in blue) with a linearized TAR vector containing a yeast selectable marker HIS3, centromeric sequence from chromosome 6 (CEN6) and two unique targeting sequences (hook1 and hook2) homologous to 5' and 3' ends of the targeted region. After co-transformation into yeast Saccharomyces cerevisiae, recombination between targeting sequences in the vector and the targeted sequences of the genomic DNA fragment leads to the rescue of the fragment as a circular YAC (yeast artificial chromosome) molecule. For TAR cloning experiments, the vector DNA is linearized by a unique endonuclease located between the hooks to expose targeting sequences.

Credit

Natalay Kouprina, email: kouprinn@mail.nih.gov

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