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Na rekombinaciji temelječa umetna evolucija kvasovke Saccharomyces cerevisiae za acidotoleranco
ID Slokar, Dejan (Author), ID Petrovič, Uroš (Mentor) More about this mentor... This link opens in a new window

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Abstract
Rast pri nizkih vrednostih pH je pri kvasovki Saccharomyces cerevisiae v industrijskem okolju dobrodošla lastnost, saj poleg pozitivnega vpliva na mnoge metabolne procese, zmanjšuje tudi možnost bakterijske kontaminacije. Acidotoleranca je poligenska lastnost, nanjo vpliva večje število genov in je zato kompleksna za preučevanje. Z namenom pridobitve na nizke vrednosti pH odpornejši sev smo iterativno križali 6 genetsko različnih sevov kvasovke S. cerevisiae. Po vsakem križanju smo s selekcijo izbrali najodpornejšo segreganto generacije, tako imenovano zmagovalko. To segreganto smo uporabili za naslednje križanje. Zmagovalka zadnje generacije je bila zmožna boljše rasti pri nižjih vrednostih pH kot vsi starševski sevi in vse ostale zmagovalke. Tako smo z iterativnim križanjem in selekcijskim postopkom uspešno izboljšali fenotip acidotolerance. Za genetsko karakterizacijo smo zmagovalko zadnje generacije za potrebe identifikacije lokusov kvantitativne lastnosti (QTL) križali s sevom Y7092 in pripravili veliko število segregant. Segregante so v tem selekcioniranem zbiru rastle v selekciji v nizkih vrednostih pH, v povprečnem zbiru pa brez selekcije. Po sekvenciranju DNA iz obeh zbirov smo z bioinformatsko analizo primerjali frekvence variacij posameznih nukleotidov v povprečnem in selekcioniranem zbiru. Določili smo 45 potencialnih QTL, znotraj njih pa predlagali 91 verjetnih vzročnih genov za acidotoleranco.

Language:Slovenian
Keywords:Saccharomyces cerevisiae, acidotoleranca, umetna evolucija, poligenske lastnosti, X-QTL, analiza QTL, iterativno križanje, izboljševanje fenotipov kvasovke
Work type:Master's thesis/paper
Organization:BF - Biotechnical Faculty
Year:2020
PID:20.500.12556/RUL-122377 This link opens in a new window
COBISS.SI-ID:44400131 This link opens in a new window
Publication date in RUL:06.12.2020
Views:892
Downloads:198
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Secondary language

Language:English
Title:Recombination-based artificial evolution of Saccharomyces cerevisiae for thermotolarance and acidotolerance
Abstract:
Ability to grow at low pH is a desirable feature for the yeast Saccharomyces cerevisiae in an industrial environment, because low pH, among many positive effects on cell metabolism, diminishes bacterial contamination. Acidotolerance is a polygenic trait, which means that there are multiple genes determining this trait. Therefore, such traits are relatively difficult to study. With the aim to obtain acidotolerant yeast strains we iteratively crossed 6 genetically different yeast strains. After each crossing we applied selection to the segregants and selected the most resistant segregant of the generation, the so-called winner. The winner of a generation was the segregant we used in the next crossing. The winner of the last generation was able to grow at lower pH then all the parental strains and all other generation winners. Thus, we have successfully improved acidotolarence yeast phenotype with iterative crossing and selection. For the identification of quantitative trait loci (QTLs), we crossed the winner of the last generation with the strain Y7092 and prepared an extremely large pool of segregants. The segregants in the extreme pool grew in selective medium with low pH, while the segregants in the average pool grew in a medium without selection. After sequencing of the pools’ DNA, we compared frequencies of single nucleotide variations between the extreme and the average pool using bioinformatics analysis. We predicted 45 potential QTLs and suggested 91 plausible causative genes for acidotolarance.

Keywords:Saccharomyces cerevisiae, acidotolerance, artificial evolution, polygenic traits, X-QTL, QTL analysis, iterative crossing, improving yeast phenotype

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