Selection of yeast cells with a higher plasmid copy number in a Saccharomyces cerevisiae autoselection system.
Autoselection systems allow the selection of a genetically engineered population independently of the growth medium composition. The structure of a Saccharomyces cerevisiae population transformed with an autoselection plasmid, in which a carbon-source-dependent modulation of the plasmid copy number occurs, was analysed. By means of flow cytometric procedures we tested the cell viability, dynamics of growth and heterologous protein production at single cell level. Such analyses allow the identification and the tracking of a specific cellular sub-population with a higher plasmid copy number which arises after the carbon source shift. The effects of the cellular plasmid distribution on the dynamics of growth are also discussed.
Compagno
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Dipartimento di Fisiologia e Biochimica Generali, Sez. Biochimica Comparata, Universita di Milano, Italy.
Porro
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D
Radice
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S
Martegani
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E
Ranzi
B M
BM
D005434:Flow Cytometry; D010957:Plasmids; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014170:Transformation, Genetic