Raugu genoma inženierija ade2Δ celmu iegūšanai un industriālās kultivēšanas potenciāla izpēte

Abstract

Bakalaura darba ietvaros veiktas raugu transformācijas ar mērķi izslēgt ADE2 gēnu. Izstrādāti DNS fragmenti raugu Kluyveromyces marxianus, Komagataella phaffii, Yarrowia lipolytica, Schizosaccharomyces pombe transformācijai ar CRISPR/Cas9 sistēmu. Rauga Saccharomyces cerevisiae transformācijai izmantotas divas metodes – transformācija ar lineāru fragmentu un CRISPR/Cas9. Saccharomyces cerevisiae transformācijām izmantoti jau laboratorijā esoši DNS fragmenti. Dažādi Saccharomyces cerevisiae adenīna auksotrofie celmi audzēti industriāli potenciāli pielietotos substrātos: ābolu sulā, vīnogu sulā un iesala ekstraktā. Veikts raugu genotipu S288C, CEN.PK, W303 prototrofo un adenīna auksotrofo celmu fermentācijas salīdzinājums iesala ekstraktā.

Within the scope of this bachelor’s thesis, yeast transformations were performed with the aim of inactivating the ADE2 gene. DNA fragments were developed for the transformation of yeasts Kluyveromyces marxianus, Komagataella phaffii, Yarrowia lipolytica, Schizosaccharomyces pombe with the CRISPR/Cas9 system. For the yeast Saccharomyces cerevisiae two methods of transformation were used – transformation with a linear fragment and CRISPR/Cas9. DNA fragments used in the transformations of Saccharomyces cerevisiae were already available in the laboratory. Various adenine auxotrophic strains of Saccharomyces cerevisiae were grown in potentially industrially used substrates: apple juice, grape juice and malt extract. A comparison of the fermentation of prototrophic and adenine auxotrophic strains of yeast genotypes S288C, CEN.PK, W303 in malt extract was performed.