Analiza uporabnosti medgenske regije 16S-23S rDNA za neposredno identifikacijo vrst ocetnokislinskih bakterij iz mikrobiot kisa in kombuče s tehnologijo sekvenciranja Illumina : magistrsko delo

Ribič, Alja

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Title:	Analiza uporabnosti medgenske regije 16S-23S rDNA za neposredno identifikacijo vrst ocetnokislinskih bakterij iz mikrobiot kisa in kombuče s tehnologijo sekvenciranja Illumina : magistrsko delo
Authors:	ID Ribič, Alja (Author) ID Trček, Janja (Mentor) More about this mentor...
Files:	MAG_Ribic_Alja_2024.pdf (912,66 KB) MD5: 8FA44717E7DD722BFA940C3C6BD90133
Language:	Slovenian
Work type:	Master's thesis/paper
Typology:	2.09 - Master's Thesis
Organization:	FNM - Faculty of Natural Sciences and Mathematics
Abstract:	V magistrskem delu smo razvili metodološki pristop za neposredno identifikacijo vrst ocetnokislinskih bakterij iz kisa in kombuče na osnovi sekvenciranja odseka medgenske regije 16S-23S rDNA s tehnologijo Illumina. Do sedaj uporabljena tarčna molekula, to je gen za 16S rRNA, je namreč v primeru ocetnokislinskih bakterij evolucijsko zelo ohranjena, zato zagotavlja zanesljivo identifikacijo le do nivoja rodu. S tem namenom smo konstruirali dva para začetnih oligonukleotidov za pomnoževanje polovice medgenske regije 16S-23S rDNA, in ju uporabili za konstrukcijo knjižnic in sekvenciranje s tehnologijo Illumina. Z obema paroma začetnih oligonukleotidov smo najprej analizirali relativno abundanco vrst ocetnokislinskih bakterij v laboratorijsko pripravljenih bakterijskih združbah. Par začetnih oligonukleotidov, ki je omogočil boljšo oceno vrstne sestave bakterijske združbe, smo nadalje uporabili za analizo mikrobiot jabolčnega, bezgovega in vinskega kisa ter napitka kombuča. Preiskani vzorci so bili proizvedeni s tradicionalnim statičnim in submerznim postopkom. Postavljena metoda je uporabna za hitro identifikacijo vrst ocetnokislinskih bakterij iz substratov, kjer te bakterije predstavljajo prevladujoči del bakterijske združbe.
Keywords:	ocetnokislinske bakterije, medgenska regija 16S-23S rDNA, Illumina, kis, kombuča
Place of publishing:	Maribor
Place of performance:	Maribor
Publisher:	[A. Ribič]
Year of publishing:	2024
Number of pages:	VII, 57 f.
PID:	20.500.12556/DKUM-90783
UDC:	579.66(043.2)
COBISS.SI-ID:	210585859
Publication date in DKUM:	08.10.2024
Views:	167
Downloads:	49
Metadata:
Categories:	FNM
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Licences

License:	CC BY 4.0, Creative Commons Attribution 4.0 International

Link:	http://creativecommons.org/licenses/by/4.0/
Description:	This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.
Licensing start date:	23.09.2024

Secondary language

Language:	English
Title:	Analysis of the Utility of 16S-23S rDNA Spacer Region for the Direct Identification of Acetic Acid Bacteria Species from the Microbiota of Vinegar and Kombucha Using Illumina Sequencing Technology
Abstract:	In the master's thesis, we developed a methodological approach for the direct identification of acetic acid bacteria species from the microbiota of vinegar and kombucha based on sequencing 16S-23S rDNA internal transcribed spacer, using Illumina technology. The previously used target molecule, the 16S rRNA gene, is evolutionarily highly conserved in acetic acid bacteria, thus providing reliable identification only up to the genus level. For this purpose, we constructed two pairs of primers to amplify half of the 16S-23S rDNA internal transcribed spacer, which were then used to construct libraries and perform sequencing using Illumina technology. These primer pairs were used to analyze the relative abundance of acetic acid bacteria species in laboratory-prepared bacterial communities. The primer pair that provided a more accurate estimation of the species composition was subsequently used to analyze microbiota of apple, elderflower, and wine vinegars, and kombucha beverage. The examined samples were produced by traditional static and submerged methods. The established method is suitable for the rapid identification of acetic acid bacteria species from substrates where these bacteria represent the dominant part of the bacterial community.
Keywords:	acetic acid bacteria, 16S-23S rDNA internal transcribed spacer, Illumina, vinegar, kombucha

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