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Mutants of Saccharomyces cerevisiae Partially Defective in the Last Steps of the Haem Biosynthetic Pathway: Isolation and Genetical Characterization

Anna Kurlandzka, J. RYTKA

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Abstract

A novel method for the isolation of Saccharomyces cerevisiae mutants partially defective in haem synthesis is described. Mutant clones were identified by their fluorescence under UV light due to the accumulation of porphyrins in cells, and by their ability to grow on nonfermentable carbon sources due to their preserved haemoprotein synthesis. Thirteen such mutants were obtained by this procedure. The defects in haem synthesis and accumulation of porphyrins in all the mutants were confirmed by spectrophotometric analysis. Complementation tests with biochemically defined, haem-less strains showed that in seven mutants uroporphyrinogen decarboxylase was affected and that in three mutants the defect concerned ferrochelatase. The defects in the remaining three mutants were not defined.

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A novel method for the isolation of Saccharomyces cerevisiae mutants partially defective in haem synthesis is described. Mutant clones were identified by their fluorescence under UV light due to the accumulation of porphyrins in cells, and by their ability to grow on nonfermentable carbon sources due to their preserved haemoprotein synthesis. Thirteen such mutants were obtained by this procedure. The defects in haem synthesis and accumulation of porphyrins in all the mutants were confirmed by spectrophotometric analysis. Complementation tests with biochemically defined, haem-less strains showed that in seven mutants uroporphyrinogen decarboxylase was affected and that in three mutants the defect concerned ferrochelatase. The defects in the remaining three mutants were not defined.

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Available abstract

A novel method for the isolation of Saccharomyces cerevisiae mutants partially defective in haem synthesis is described. Mutant clones were identified by their fluorescence under UV light due to the accumulation of porphyrins in cells, and by their ability to grow on nonfermentable carbon sources due to their preserved haemoprotein synthesis. Thirteen such mutants were obtained by this procedure. The defects in haem synthesis and accumulation of porphyrins in all the mutants were confirmed by spectrophotometric analysis. Complementation tests with biochemically defined, haem-less strains showed that in seven mutants uroporphyrinogen decarboxylase was affected and that in three mutants the defect concerned ferrochelatase. The defects in the remaining three mutants were not defined.

Key concepts: Saccharomyces cerevisiae, Mutant, Isolation (microbiology), Biochemistry, Biology, Genetics, Chemistry, Yeast

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