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- W2073249737 abstract "A general method to replace chromosomal DNA sequences of Saccharomyces cerevisiae by any in vitro modified DNA sequence has been developed and was applied to the PHO5 locus on chromosome II. A recipient strain was constructed in which part of the chromosomal PHO5 sequence was substituted by the URA3 gene. Replacement of this pho5-URA3 substitution by pho5 mutant alleles was achieved in one step by cotransformation with a pho5 DNA fragment and the self-replicating plasmid YEp13, which contains the LEU2 gene as a selectable marker. Leu+ transformants were selected, and the replacement events at the PHO5 locus were detected by their Ura− phenotype (1–4 % of the Leu+ were Ura−). In a similar way the PHO5 coding sequence was replaced by the sequence coding for human tissue-type plasminogen activator (t-PA)." @default.
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- W2073249737 date "1985-01-01" @default.
- W2073249737 modified "2023-10-14" @default.
- W2073249737 title "One-step gene replacement in yeast by cotransformation" @default.
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- W2073249737 doi "https://doi.org/10.1016/0378-1119(85)90072-1" @default.
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