Abstract
This chapter describes the preparation of cytochrome-deficient mutants of yeast. The types of mutants considered in two genetic classes are: (1) the cytoplasmic or ρ– mutants (vegetative petities) and (2) the chromosomal mutants (segregational mutants), which can be further classified as p or cy. Different chromosomal mutants can have a variety of partial or complete deficiencies. It is also possible to prepare mutant strains having more than one genetic defect, and thereby increase the types of cytochrome deficiencies. For example, chromosomal mutants (p and cy) usually can be obtained both as ρ+ and ρ–. Haploid strains can be constructed by genetic recombination to have two or more mutant genes. Any mutagenic treatment, which is used to produce general types of chromosomal mutants, also can be employed to obtain cytochrome mutants. Ultraviolet irradiation and the chemical mutagens nitrous acid and nitrosoguanidine (N-methyl-N'-nitro-N-nitrosoguanidine) have been found to be effective. It is possible to obtain yeast mutants that have defects in the tricarboxylic acid cycler and mutants, which have high concentrations of porphyrins and metalloporphyrins. The chapter also discusses the preparation of ρ– strains, the preparation of chromosomal mutants, isolation procedures, and types of mutants.
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