Hybridization of Candida albicans through fusion of protoplasts |
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Authors: | Alvin Sarachek Douglas D Rhoads Robert H Schwarzhoff |
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Institution: | (1) Department of Biological Sciences, Wichita State University, 67208 Wichita, Kansas, USA |
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Abstract: | Protoplasts of complementing auxotrophs of Candida albicans can fuse in the presence of polyethylene glycol and generate prototrophic cells. The yields of prototrophs from fusion mixtures
depend greatly on the particular combinations of auxotrophies involved but not on other features of the strain backgrounds
of protoplasts. The initial cellular products of fusions isolated on selective media are heterokaryons which replicate slowly
but also segregate single parental nuclei into blastospores in high frequency. Karyogamy within heterokaryons produces hybrid
nuclei which, on segregation, give rise to rapidly growing, uninucleate substrains. Analyses of the substrains show that hybrid
nuclei either stabilize as diploid or undergo random loss of chromosomes to stabilize at various levels of aneuploidy prior
to segregation. Chromosome losses and radiation induced mitotic crossing-over can effect recombination for parental auxotrophic
markers in hybrids; patterns of recombination for ader and arg markers provide the first documented example of chromosomal linkage in C. albicans. Thus, protoplast fusions offer opportunities otherwise unavailable for applying the incisive tools of genetic recombination
to analysis of this important, asexual yeast. |
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Keywords: | Candida albicans Heterokaryons Mitotic recombination Protoplast fusion Hybrids |
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