prikaz prve stranice dokumenta Analiza ekspresije icl gena u Mycobacterium smegmatis pomoću time-lapse fluorescentne mikroskopije
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master's thesis
Analiza ekspresije icl gena u Mycobacterium smegmatis pomoću time-lapse fluorescentne mikroskopije
2016. urn:nbn:hr:193:083599
Fabijanić, Martina
University of Rijeka
Faculty of Biotechnology and Drug Development

Cite this document

Fabijanić, M. (2016). Analiza ekspresije icl gena u Mycobacterium smegmatis pomoću time-lapse fluorescentne mikroskopije (Master's thesis). Rijeka: University of Rijeka. Retrieved from https://urn.nsk.hr/urn:nbn:hr:193:083599

Fabijanić, Martina. "Analiza ekspresije icl gena u Mycobacterium smegmatis pomoću time-lapse fluorescentne mikroskopije." Master's thesis, University of Rijeka, 2016. https://urn.nsk.hr/urn:nbn:hr:193:083599

Fabijanić, Martina. "Analiza ekspresije icl gena u Mycobacterium smegmatis pomoću time-lapse fluorescentne mikroskopije." Master's thesis, University of Rijeka, 2016. https://urn.nsk.hr/urn:nbn:hr:193:083599

Fabijanić, M. (2016). 'Analiza ekspresije icl gena u Mycobacterium smegmatis pomoću time-lapse fluorescentne mikroskopije', Master's thesis, University of Rijeka, accessed 07 December 2024, https://urn.nsk.hr/urn:nbn:hr:193:083599

Fabijanić M. Analiza ekspresije icl gena u Mycobacterium smegmatis pomoću time-lapse fluorescentne mikroskopije [Master's thesis]. Rijeka: University of Rijeka; 2016 [cited 2024 December 07] Available at: https://urn.nsk.hr/urn:nbn:hr:193:083599

M. Fabijanić, "Analiza ekspresije icl gena u Mycobacterium smegmatis pomoću time-lapse fluorescentne mikroskopije", Master's thesis, University of Rijeka, Rijeka, 2016. Available at: https://urn.nsk.hr/urn:nbn:hr:193:083599

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Metadata
TitleAnaliza ekspresije icl gena u Mycobacterium smegmatis pomoću time-lapse fluorescentne mikroskopije
Title (english)Analysis of icl expression in Mycobacterium smegmatis by using fluorescent time-lapse microscopy
AuthorMartina Fabijanić
MentorŽeljka Maglica (mentor)
Committee memberŽeljka Maglica (predsjednik povjerenstva)
Committee memberMiranda Mladinić Pejatović (član povjerenstva)
Committee memberJasminka Giacometti (član povjerenstva)
GranterUniversity of Rijeka
(Faculty of Biotechnology and Drug Development)
Rijeka
Defense date and country2016-10-28, Croatia
Scientific / art field,
discipline and subdiscipline		BIOTECHNICAL SCIENCES
Biotechnology
Abstract
Mycobacterium smegmatis se, za razliku od M. tuberculosis, odlikuje brzim rastom i nepatogeno..u .to ga .ini prikladnim za laboratorijska ispitivanja. Ustanovljena va.nost icl gena za patogenost, virulenciju i pre.ivljavanje M. tuberculosis u .ivom organizmu potaknula je prou.avanje ekspresije icl gena M. smegmatis prilikom kultivacije na acetatu, supstratu koji u tijelu doma.ina nastaje ƒŔ-oksidacijom lipida te ulazi u anaplerotski glioksilatni ciklus. Djelovanje glioksilatnog ciklusa posredovano je djelovanjem enzima izocitrat liaze (ICL) kojeg kodiraju icl1 i icl2 geni. Metoda time-lapse fluorescentne mikroskopije, uz obradu filmova u BactImAS platformi omogu.ila je prou.avanje dinamike ekspresije icl gena u realnom vremenu (eng. real-time). Pokazana je uskla.enost kolonija, ali i individualnost pojedina.nih stanica. Inicijalni val ekspresije odvijao se istovremeno kod svih bakterija u koloniji, samo 2 sata nakon dodatka acetata. Nakon toga, svaka stanica uspostavila je vlastitu dinamiku ekspresije koja se kod mnogih stanica poja.ala neposredno prije diobe. Metaboli.ki ciklusi koje stanice koriste za pre.ivljavanje na acetatu u po.etku podupiru diobu, ali ne i rast jer je stopa rasta opadala neovisno o razini ekspresije icl gena. Povr.ina sestrinskih stanica razlikovala se vi.e od 1.5 puta u pribli.no 50% slu.ajeva, kao i ukupna fluorescencija (eng. total fluorescence). S druge strane, srednja fluorescencija (eng. mean fluorescence) je kod ve.ine sestrinskih stanica bila podjednaka. U radu je za analizu ekspresije icl gena kori.ten BactImAS softver za obradu i analizu time-lapse filmova, unaprije.ena je funkcija pripreme filogenetskih stabala i specificirane su karakteristike filmova prikladnih za obradu u programu. Podaci dobiveni iz filmova pomo.u BactImAS platforme postavljaju temelje za mnogobrojne analize u budućnosti.
Abstract (english)
Due to its fast growth and non-pathogenic nature, Mycobacterium smegmatis is a convenient experimental model for research of its close cousin, M. tuberculosis. The established relevance of icl genes for M. tuberculosis virulence, pathogenicity and survival in host organisms has encouraged icl gene expression analysis in M. smegmatis during cultivation on acetate, a substrate that is derived from β-oxidation of host lipids after which it enters the anaplerotic glyoxylate cycle. The enzyme isocytrate lyase (ICL), which is coded by icl1 and icl2 genes, participates in the glyoxylate cycle. Time-lapse fluorescent microscopy, along with movie processing in BactImAS software, has enabled analysis of icl gene expression dynamics through time. Here, the individuality of each cell is shown along with synchronicity of bacterial colonies. The initial wave of expression occurs only 2 hours after acetate addition in all cells of the colony. After that, each cell establishes an individual dynamic of expression which is enhanced in most cells before cell division occurs. The growth rate was decreasing despite the level of icl expression so it was concluded that the anaplerotic pathways which enable acetate utilization are used primarily for the establishment of cell divison, not cell growth. The area of sister cells had more than 1.5 time difference in almost 50% of cases, as did the total fluorescence. On the other hand, mean fluorescence values were predominantly equal among sister cells. Working on gene expression analysis also helped to improve BactImAS software for time-lapse movie processing and analysis. The BTree function was enhanced and movie characteristics that are compatible for BactImAS were specified. The amount of data extracted with the use of BactImAS has set a foundation for comprehensive future analysis.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:193:083599
Study programmeTitle: Biotechnology in medicine
Study programme type: university
Study level: graduate
Academic / professional title: magistar/magistra biotehnologije u medicini (magistar/magistra biotehnologije u medicini)
Type of resourceText
File originBorn digital
Access conditionsClosed access
Terms of useLicence In copyright icon
Created on2016-11-16 11:40:12