Complementation analysis with new genetic markers in Phaffia rhodozyma |
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Authors: | P Retamales R León C Martínez G Hermosilla G Pincheira V Cifuentes |
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Institution: | (1) Laboratorio de Genética, Facultad de Ciencias, Universidad de Chile, Casilla 653, Santiago, Chile |
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Abstract: | Isolation and characterization of auxotrophic mutants from wild-type and astaxanthin mutant strains of Phaffia rhodozyma is described. Differences in survival were observed when u.v. irradiation of P. rhodozyma wild-type and astaxanthin mutant strains were incubated in the dark or exposed to photoreactivating light. Ultra-violet mutagenesis was not effective to produce auxotrophic mutants in this yeast. Auxotrophic mutants were obtained with high efficiency through a nystatin enrichment procedure after a N-methyl-N-nitro-N-nitrosoguanidine (NTG) mutagenic treatment with a 0.12% survivor level. Stringent mutagenetic conditions were needed to obtain P. rhodozyma auxotrophs. The most frequent mutants were ade- and met- in a rather narrow auxotroph spectrum. These results may be associated with a possible diploid condition of this yeast. The high number of adenine auxotrophs obtained in relation to other auxotrophic mutants suggests the possibility of some degree of heterozygosity in the wild-type strain UCD 67-385. |
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Keywords: | auxotrophs mutagenesis nystatin enrichment rotoplasts fusion Phaffia rhodozyma |
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