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The Green Monster Process for the Generation of Yeast Strains Carrying Multiple Gene Deletions
Published on: December 15, 2012
Recovery of gene function by gene duplication in Saccharomyces cerevisiae
M L Bach1, F Roelants, J De Montigny
1Laboratoire de Microbiologie et Génétique, URA 1481 Université Louis Pasteur/CNRS, Strasbourg, France.
Abstract:
A prototroph revertant (Rev9) selected from an ATCase- mutant of the URA2 gene containing three nonsense mutations was shown to contain two ATCase coding sequences. We cloned both ATCase coding areas to show that the duplicated locus (dl9) was the only functional one. Its size corresponded roughly to the second half of the URA2 wild-type gene. Sequence analysis of the 5' end of dl9 indicated that this duplicated sequence was inserted within the intergenic region close to the MRS3 gene and was transcribed from an unknown promoter divergently from the MRS3 gene. The event leading to the revertant strain Rev9 included a rearrangement that increased the size of chromosome X by about 60 kb. In agreement with such a rearrangement, recombination was undetectable in the vicinity of the locus dl9. Genetic mapping confirms that the MRS3 gene is 2 cM distal to the URA2 gene on the right arm of chromosome X.
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