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master's thesis
Utjecaj brzine rasta na efikasnost adaptacije sustavom CRISPR-Cas u bakterija Escherichia coli različitih genetičkih pozadina
Zagreb: University of Zagreb, Faculty of Science, 2022. urn:nbn:hr:217:188517
Knechtl, Patricija
University of Zagreb
Faculty of Science
Department of Biology

Institutional repository: Repository of the Faculty of Science

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Knechtl, P. (2022). Utjecaj brzine rasta na efikasnost adaptacije sustavom CRISPR-Cas u bakterija Escherichia coli različitih genetičkih pozadina (Master's thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:188517

Knechtl, Patricija. "Utjecaj brzine rasta na efikasnost adaptacije sustavom CRISPR-Cas u bakterija Escherichia coli različitih genetičkih pozadina." Master's thesis, University of Zagreb, Faculty of Science, 2022. https://urn.nsk.hr/urn:nbn:hr:217:188517

Knechtl, Patricija. "Utjecaj brzine rasta na efikasnost adaptacije sustavom CRISPR-Cas u bakterija Escherichia coli različitih genetičkih pozadina." Master's thesis, University of Zagreb, Faculty of Science, 2022. https://urn.nsk.hr/urn:nbn:hr:217:188517

Knechtl, P. (2022). 'Utjecaj brzine rasta na efikasnost adaptacije sustavom CRISPR-Cas u bakterija Escherichia coli različitih genetičkih pozadina', Master's thesis, University of Zagreb, Faculty of Science, accessed 17 May 2024, https://urn.nsk.hr/urn:nbn:hr:217:188517

Knechtl P. Utjecaj brzine rasta na efikasnost adaptacije sustavom CRISPR-Cas u bakterija Escherichia coli različitih genetičkih pozadina [Master's thesis]. Zagreb: University of Zagreb, Faculty of Science; 2022 [cited 2024 May 17] Available at: https://urn.nsk.hr/urn:nbn:hr:217:188517

P. Knechtl, "Utjecaj brzine rasta na efikasnost adaptacije sustavom CRISPR-Cas u bakterija Escherichia coli različitih genetičkih pozadina", Master's thesis, University of Zagreb, Faculty of Science, Zagreb, 2022. Available at: https://urn.nsk.hr/urn:nbn:hr:217:188517

Metadata
TitleUtjecaj brzine rasta na efikasnost adaptacije sustavom CRISPR-Cas u bakterija Escherichia coli različitih genetičkih pozadina
Title (english)The influence of growth rate on the efficiency of the CRISPR-Cas mediated adaptation in Escherichia coli of different genetic backgrounds
AuthorPatricija Knechtl
MentorIvana Ivančić Baće (mentor)
MentorDamjan Mitić (komentor)
Committee memberIvana Ivančić Baće (predsjednik povjerenstva)
Committee memberMirela Sertić Perić (član povjerenstva)
Committee memberDraginja Mrvoš-Sermek (član povjerenstva)
Committee memberŽeljka Soldin (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Science
(Department of Biology)
Zagreb
Defense date and country2022-10-25, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Biology
Abstract
Sustav CRISPR-Cas sastoji se od lokusa CRISPR koji zajedno s pridruženim genima cas omogućuje adaptivnu imunost mnogim bakterijama i većini arheja protiv stranih genetičkih elemenata. Lokus CRISPR sastoji se od brojnih ponavljajućih sekvenci DNA koje su razdvojene kratkim razmaknicama strane DNA. Naivna adaptacija prva je faza zaštite u kojoj se kratki fragmenti iz stranih organizama ugrađuju u lokus CRISPR kako bi se zaštitila stanica od budućih napada istom stranom DNA. U ovom radu istražen je utjecaj ekspresije gena cas1 i cas2 na brzinu rasta stanica, određena je faza rasta u kojoj dolazi do ugradnje nove razmaknice te je istražena efikasnost adaptacije u tri različita bakterijska soja E. coli koji imaju tri različite genetičke pozadine (BW25113, AB1157 i MG1655). Također, istraženo je kako količine proteina Cas1 i Cas2 utječu na efikasnost naivne adaptacije (ugradnju razmaknice). Na temelju analize dobivenih rezultata može se zaključiti kako se usporeni rast bakterijskih stanica javlja zbog ugradnje razmaknice u lokus CRISPR, u ranoj log fazi rasta, samo u genetičkoj pozadini BW25113. U bakterijskim sojevima koji su rasli sporo došlo je do ekspresije proteina Cas1 i Cas2, dok geni rpoS i crl nemaju ulogu u izgradnji adaptivne imunosti procesom naivne adaptacije.
Abstract (english)
The CRISPR-Cas system consists of the CRISPR locus, which, together with the CRISPRassociated (cas) genes, provides adaptive immunity to many bacteria and most archaea against foreign genetic elements. The CRISPR locus consists of numerous repetitive DNA sequences separated by short spacers that are derived from foreign DNA. Naïve adaptation is the first phase of protection in which short fragments from foreign organisms are incorporated into the CRISPR locus to protect the cell from future attacks by the same foreign DNA. In this research, we investigated the influence of cas1 and cas2 gene expression on cell growth rate, we determined in which growth phase a new spacer is incorporated and we investigated the efficiency of adaptation in three different bacterial strains of E. coli with three different genetic backgrounds (BW25113, AB1157 and MG1655). Also, we investigated how the amounts of Cas1 and Cas2 proteins affect the efficiency of adaptation (spacer acquisition). Based on the analysis of the obtained results, it can be concluded that the slowed growth of bacterial cells occurs due to the insertion of a spacer into the CRISPR locus in the early log phase of growth only in the BW25113 genetic background. In bacterial strains that grew slowly, Cas1 and Cas2 proteins were expressed. Genes rpoS and crl have no role in building CRISPR immunity through the process of naïve adaptation.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:217:188517
Study programmeTitle: Integrated undergraduate and graduate university study of Biology and Chemistry Education; specializations in: Integrated undergraduate and graduate university study of Biology and Chemistry Education
Course: Integrated undergraduate and graduate university study of Biology and Chemistry Education
Study programme type: university
Study level: integrated undergraduate and graduate
Academic / professional title: magistar/magistra edukacije biologije i kemije (magistar/magistra edukacije biologije i kemije)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Embargo expiration date: 2023-08-25
Terms of useLicence In copyright icon
Created on2022-11-03 13:08:47