Interaction between nucleus and mitochondria in reconstituted cells.

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In order to study the interaction between nucleus and mitochondria in yeasts,we tried to develop in vitro cell system which reconstituted from anucleated cell, nucleus and mitochondria. For this purpose, we carried out the following investigations.1. Isolation of mutants which is suitable for cell reconstitution and transformation.2. Establishment of protplast conversion and its regeneration in giant yeasts,Wickerhamia fluorescene and Candida ingens.3. Development of versatile apparatus for pulsed field gel electrophoresis to separate large nuclear DNA, mtDNA and plasmid DNA. Determination of electrophoretic karyotypes of various yeasts by the device.4. Development of yeast genetic analysis programs (YGAP) for a microcomputer.5. Introduction of stepwise transformation which consists of transformation of conventional host cells and subsequent cytoductive transfer of vectors to destination cells with mitochondria by using kar1 mutation.6. Induction of heterothallic strains from homothallic strains by mutation and subseqent genetic analysis of mitochondria by mating.