Secuenciamiento de levadura oleaginosa Rhodotorula mucilaginosa P11
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Date
2023
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Universidad Nacional de Trujillo
Abstract
Una alternativa de solución hacia el impacto ambiental debido al uso de tierras agrícolas para la elaboración de oleoquímicos, es el uso de levaduras oleaginosas productoras de lípidos en más del 25% de su peso seco, los cuales están formados por ácidos grasos insaturados. El propósito de esta tesis fue secuenciar la levadura oleaginosa Rhodotorula mucilaginosa P11 que presenta un porcentaje de lípidos de 67.4% en base seca. Para alcanzar éste objetivo se produjo biomasa en medio YPD (30 °C/150 rpm/10 h), seguidamente se extrajo el ADN genómico mediante el método tradicional utilizando cloroformo y se determinó su concentración y calidad mediante espectrofotometría Nanodrop 2000c y electroforesis en gel de agarosa 1.2%. El ADN genómico fue secuenciado utilizando la tecnología Illumina Miseq pair-end 2x150 pb y se realizó un control de calidad a los datos obtenidos usando el software FastQC, obteniéndose un total de 6 308 858 secuencias forward y 6 308 858 secuencias reverse con un %GC de 56%. Estas secuencias obtenidas van a permitir realizar posteriormente un ensamblaje y con ello conocer sus características metabólicas y funcionales para su posterior análisis.
ABSTRACT An alternative solution to the environmental impact due to the use of agricultural land to produce oleochemicals, is the use of oleaginous yeasts that produce of accumulating lipids in more than 25% of their dry weight, which are formed by unsaturated fatty acids. The aim of this thesis was to sequence the oleaginous yeast Rhodotorula mucilaginosa P11 that presents a lipid percentage of 67.4% on a dry basis. To reach this aim, biomass was produced in YPD medium (30 °C/ 150 rpm/ 10 hours), followed by genomic DNA extraction using the traditional chloroform method. The DNA concentration and its quality were determined using Nanodrop 2000c spectrophotometry and 1.2% agarose gel electrophoresis. The genomic DNA was sequenced using Illumina Miseq pair-end 2x150 bp technology and a quality control was performed on the data obtained using the FastQC software, obtaining a total of 6,308,858 forward sequences and 6,308,858 reverse sequences with a %GC of 56%. These sequences obtained will allow a subsequent assembly and thus to know their metabolic and functional characteristics for further analysis.
ABSTRACT An alternative solution to the environmental impact due to the use of agricultural land to produce oleochemicals, is the use of oleaginous yeasts that produce of accumulating lipids in more than 25% of their dry weight, which are formed by unsaturated fatty acids. The aim of this thesis was to sequence the oleaginous yeast Rhodotorula mucilaginosa P11 that presents a lipid percentage of 67.4% on a dry basis. To reach this aim, biomass was produced in YPD medium (30 °C/ 150 rpm/ 10 hours), followed by genomic DNA extraction using the traditional chloroform method. The DNA concentration and its quality were determined using Nanodrop 2000c spectrophotometry and 1.2% agarose gel electrophoresis. The genomic DNA was sequenced using Illumina Miseq pair-end 2x150 bp technology and a quality control was performed on the data obtained using the FastQC software, obtaining a total of 6,308,858 forward sequences and 6,308,858 reverse sequences with a %GC of 56%. These sequences obtained will allow a subsequent assembly and thus to know their metabolic and functional characteristics for further analysis.
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Keywords
Illumina, Rhodotorula mucilaginosa, Levadura oleaginosa, Secuenciamiento