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Classic techniques for improvement of industrial yeast strains: Part III ? A method for enucleation of Saccharomyces sp.

100%

Salek A. T.

Biotechnologia

2002

nr 1

187-194

The first method for enucleation of yeast Saccharomyces cerevisiae is reported. Various strains, including some killer strain and respiratory-deficient mutants of Saccharomyces cerevisiae were enucleated after treatment with cytochalasin B. Removal of nuclei from protruding sphaeroplasts was induced by centrifugation in a Percoll density gradient. The enucleation yield (which averaged about 80%) and the quality of the cytoplasts were best when the yeast culture had been synchronized with nocodazole before the preparation. The presence of 1 mM CaCl2 and ATP (10 muM) in the enucleation medium prevented the formation of fragile products or aggregation. Cytoplasts could be stored for at least 1 day without visible deterioration.

The use of new strategies in the cloning studies on somatic cell cloning in pigs

75%

Skrzyszowska M. , Samiec M.

Biotechnologia

2006

nr 1

68-81

Somatic cell nuclear transfer (SCNT) technique in pigs remains relatively low (2% to 5% of produced piglets), that is why further efforts have to be made to optimize both a multi-step cloning procedure and to improve a structuro-functional quality of recipient oocytes and nuclear donor cells. Pre- and postimplantation developmental potential of porcine SCNT-derived embryos depends to a high degree on not only coordination of mitotic cycle stage with phenotype of nuclear donor cell, but also proper combination of the methods of maternal chromosome elimination (enucleation), oocyte reconstruction techniques, the systems of artificial activation of generated nuclear-cytoplasmic hybrids (clonal cybrids) and in vitro culture of reconstructed embryos. Generally, it can result in increasing the competences of both somatic nuclear and mitochondrial genome for epigenetic remodeling/reprogramming in developing cloned embryos.