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master's thesis
Ekspresija gena staničnog odgovora u mre11-2 mutantnoj liniji Arabidopsis thaliana (L.) nakon indukcije dvolančanih lomova DNA Bleomicinom
Split: University of Split, University of Split, Faculty of science, 2016. urn:nbn:hr:166:035137
Rakušić, Mia
University of Split
Faculty of Science
Department of Biology

Institutional repository: Repository of Faculty of Science

Cite this document

Rakušić, M. (2016). Ekspresija gena staničnog odgovora u mre11-2 mutantnoj liniji Arabidopsis thaliana (L.) nakon indukcije dvolančanih lomova DNA Bleomicinom (Master's thesis). Split: University of Split, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:166:035137

Rakušić, Mia. "Ekspresija gena staničnog odgovora u mre11-2 mutantnoj liniji Arabidopsis thaliana (L.) nakon indukcije dvolančanih lomova DNA Bleomicinom." Master's thesis, University of Split, Faculty of Science, 2016. https://urn.nsk.hr/urn:nbn:hr:166:035137

Rakušić, Mia. "Ekspresija gena staničnog odgovora u mre11-2 mutantnoj liniji Arabidopsis thaliana (L.) nakon indukcije dvolančanih lomova DNA Bleomicinom." Master's thesis, University of Split, Faculty of Science, 2016. https://urn.nsk.hr/urn:nbn:hr:166:035137

Rakušić, M. (2016). 'Ekspresija gena staničnog odgovora u mre11-2 mutantnoj liniji Arabidopsis thaliana (L.) nakon indukcije dvolančanih lomova DNA Bleomicinom', Master's thesis, University of Split, Faculty of Science, accessed 23 November 2024, https://urn.nsk.hr/urn:nbn:hr:166:035137

Rakušić M. Ekspresija gena staničnog odgovora u mre11-2 mutantnoj liniji Arabidopsis thaliana (L.) nakon indukcije dvolančanih lomova DNA Bleomicinom [Master's thesis]. Split: University of Split, Faculty of Science; 2016 [cited 2024 November 23] Available at: https://urn.nsk.hr/urn:nbn:hr:166:035137

M. Rakušić, "Ekspresija gena staničnog odgovora u mre11-2 mutantnoj liniji Arabidopsis thaliana (L.) nakon indukcije dvolančanih lomova DNA Bleomicinom", Master's thesis, University of Split, Faculty of Science, Split, 2016. Available at: https://urn.nsk.hr/urn:nbn:hr:166:035137

Metadata
TitleEkspresija gena staničnog odgovora u mre11-2 mutantnoj liniji Arabidopsis thaliana (L.) nakon indukcije dvolančanih lomova DNA Bleomicinom
Title (english)Expression of damage response genes in mre11-2 mutant line of Arabidopsis thaliana (L.) after inducing DNA double strand breaks with Bleomycin treatment
AuthorMia Rakušić
MentorIvica Šamanić (mentor)
MentorJasna Puizina (mentor)
Committee memberIvica Šamanić (predsjednik povjerenstva)
Committee memberJasna Puizina (član povjerenstva)
Committee memberŽeljana Fredotović (član povjerenstva)
GranterUniversity of Split
Faculty of Science
(Department of Biology)
Split
Defense date and country2016-09-22, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Biology
Abstract
Protein Mre11, u okviru kompleksa Mre11-Rad50-Nbs1/Xrs2 (MRN/X), ima važnu ulogu u popravcima dvolančanih lomova DNA svih do sada istraživanih organizama, od bakterija do čovjeka. U cilju istraživanja te specifične uloge proteina Mre11 u biljne modelne vrste Arabidopsis thaliana (L.), uspoređena je ekspresija gena MRE11 i drugih gena staničnog odgovora, nakon izazivanja dvolančanih lomova DNA bleomicinom. Usporedna analiza ekspresije ciljanih gena urađena je kod mutantne liniji mre11-2 s insercijom T-DNA u 19. egzonu i kod biljaka divljeg tipa, ekotipova Columbia [Col-0] i Wassilewskija [Ws]. Metodom RT-PCR ukupna biljna RNA prevedena je u cDNA koja je poslužila kao kalup za umnažanje gena metodom PCR. Umnoženi geni su potom naneseni na agarozni gel kako bi se mogla odrediti približna ekspresija tih gena kod različitih uzoraka. U mutanata mre11-2 tretiranih bleomicinom rezultati RT-PCR-a pokazali su smanjenu transkripciju gena CYCB1;1, biljega faze G2/M staničnog ciklusa, te gena ključnih za popravak DNA (BRCA1, RAD51, PARP1), za razliku od povećane ekspresije tih gena kod biljaka divljeg tipa. Ekotipovi Columbia i Wassilewskija pokazuju podudaran obrazac ekspresije gena staničnog odgovora. Smanjena ekspresija gena staničnog odgovora u mutantnoj liniji Atmre11-2 potvrđuju važnost cjelovitosti Mre11 proteina za normalni odgovor stanice na dvolančane lomove DNA.
Abstract (english)
Mre11 protein, a part of the MRE11-RAD50-Nbs1/Xrs2 (MRN/X) complex, plays an important role in the repair of DNA double strand breaks in all organisms studied so far, from bacteria to human. In order to investigate this specific role of MRE11 in plant model species Arabidopsis thaliana (L.), we compared the expression of MRE11 gene and other DNA damage response genes, after DSB induction by bleomycin treatment. For comparison, we performed an analogous expression analysis in the mre11-2 mutant line with T-DNA insertion in the 19th exon and in the wild type ecotypes Columbia [Col-0] and Wassilewskija [Ws]. Using RT-PCR, total plant RNAs were transformed into cDNA, which served as a template for PCR gene amplification. These genes are then loaded into an agarose gel to determine the gene expression in various samples. In bleomycin treated mre11-2 mutant, RT-PCR showed reduced transcription of G2/M-specific CYCB1;1 marker gene and key DNA damage repair genes (BRCA1, RAD51, PARP1) in contrast with the increased expression of these genes in wild plants. Columbia and Wassilewskija ecotypes showed similar expression of genes involved in cellular stress response. Expression of DNA damage response genes in the Atmre11-2 mutant line confirms the importance of full length Mre11 protein for normal cell response to DNA double strand breaks.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:166:035137
Study programmeTitle: Biology and Chemistry; specializations in: major in teaching
Course: major in teaching
Study programme type: university
Study level: graduate
Academic / professional title: magistar/magistra edukacije biologije i kemije (magistar/magistra edukacije biologije i kemije)
Type of resourceText
File originBorn digital
Access conditionsAccess restricted to students and staff of home institution
Terms of useLicence In copyright icon
Created on2017-05-19 15:31:06