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master's thesis
Određivanje količine proteina Cas3 u ovisnosti o jačini promotora, temperaturi i šaperonu HtpG u bakteriji Escherichia coli
Zagreb: University of Zagreb, Faculty of Science, 2021. urn:nbn:hr:217:291835
Barbarić, Lea
University of Zagreb
Faculty of Science
Department of Chemistry

Institutional repository: Repository of the Faculty of Science

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Barbarić, L. (2021). Određivanje količine proteina Cas3 u ovisnosti o jačini promotora, temperaturi i šaperonu HtpG u bakteriji Escherichia coli (Master's thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:291835

Barbarić, Lea. "Određivanje količine proteina Cas3 u ovisnosti o jačini promotora, temperaturi i šaperonu HtpG u bakteriji Escherichia coli." Master's thesis, University of Zagreb, Faculty of Science, 2021. https://urn.nsk.hr/urn:nbn:hr:217:291835

Barbarić, Lea. "Određivanje količine proteina Cas3 u ovisnosti o jačini promotora, temperaturi i šaperonu HtpG u bakteriji Escherichia coli." Master's thesis, University of Zagreb, Faculty of Science, 2021. https://urn.nsk.hr/urn:nbn:hr:217:291835

Barbarić, L. (2021). 'Određivanje količine proteina Cas3 u ovisnosti o jačini promotora, temperaturi i šaperonu HtpG u bakteriji Escherichia coli', Master's thesis, University of Zagreb, Faculty of Science, accessed 25 November 2024, https://urn.nsk.hr/urn:nbn:hr:217:291835

Barbarić L. Određivanje količine proteina Cas3 u ovisnosti o jačini promotora, temperaturi i šaperonu HtpG u bakteriji Escherichia coli [Master's thesis]. Zagreb: University of Zagreb, Faculty of Science; 2021 [cited 2024 November 25] Available at: https://urn.nsk.hr/urn:nbn:hr:217:291835

L. Barbarić, "Određivanje količine proteina Cas3 u ovisnosti o jačini promotora, temperaturi i šaperonu HtpG u bakteriji Escherichia coli", Master's thesis, University of Zagreb, Faculty of Science, Zagreb, 2021. Available at: https://urn.nsk.hr/urn:nbn:hr:217:291835

Metadata
TitleOdređivanje količine proteina Cas3 u ovisnosti o jačini promotora, temperaturi i šaperonu HtpG u bakteriji Escherichia coli
Title (english)Determination of the amount of Cas3 protein depending on the strenght of the promoter, temperature and chaperone HtpG in Escherichia coli
AuthorLea Barbarić
MentorIvana Ivančić Baće (mentor)
Committee memberMarko Močibob (predsjednik povjerenstva)
Committee memberIvana Ivančić Baće (član povjerenstva)
Committee memberĐani Škalamera (član povjerenstva)
Committee memberBranimir Bertoša (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Science
(Department of Chemistry)
Zagreb
Defense date and country2021-07-15, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Chemistry
Abstract
Zbog učestalih napada bakteriofaga, veliki udio bakterija razvilo je specifičan obrambeni sustav adaptivne imunosti koji se naziva CRISPR-Cas. Sustav se sastoji od seta gena cas i lokusa CRISPR koji se sastoji od kratkih ponavljajućih sljedova između kojih se ugrađuju razmaknice koje potječu od strane DNA. U bakteriji Escherichia coli efikasnost zaštite bakterija od faga sustavom CRISPR-Cas ovisi o aktivnosti proteina Cas3 koja pak ovisi o temperaturi inkubacije i šaperonu HtpG. U ovom radu istraženo je utječe li šaperon HtpG na količinu proteina Cas3 u ovisnosti o temperaturi i jačini promotora. Uz to, istraženo je da li promjene u konformaciji šaperona HtpG utječu na aktivnost proteina Cas3, odnosno na efikasnost zaštite od infekcije. Rezultati sugeriraju da šaperon HtpG utječe na funkcionalne količine proteina Cas3 u ovisnosti o temperaturi, dok točkaste mutacije u proteinu HtpG nemaju utjecaja na interakciju HtpG s proteinom Cas3. Jačina promotora za gen cas3 je ključna i analiza količina proteina Cas3 je moguća samo nakon ekspresije s inducibilnog promotora.
Abstract (english)
Due to frequent bacteriophage attacks, a large group of bacteria have developed a specific adaptive immunity defense system called CRISPR-Cas. The system consists of a set of cas genes and CRISPR locus consisting of short repeats between which spacers derived from foreign DNA are inserted. In Escherichia coli, the effectiveness of immunity against phages depends on the activity of Cas3, which depends on the incubation temperature and HtpG chaperone. In this work, the influence of HtpG chaperone on the amount of Cas3 protein as a function of temperature and promoter strength was investigated. In addition, it was investigated whether conformational changes of HtpG affect the activity of Cas3, ie the effectiveness of the immunity against infections. The results suggest that chaperone HtpG affects functional amounts of Cas3 protein as a function of temperature, whereas point mutations in HtpG protein have no effect on the interaction of HtpG with Cas3. The strength of the promoter for the cas3 gene is critical and analysis of the amount of Cas3 is only possible from an inducible promoter.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:217:291835
ProjectNumber: IP-2016-06-8861
Title: Cas3 kao kontrolna točka obrane CRISPR-Cas: razjašnjenje njegove regulacije istraživanjem stabilnosti proteina i prepisivanja u bakteriji Escherichia coli
Title: Cas3 as a key checkpoint for CRISPR-Cas immunity: Elucidating its regulation via protein stability and transcriptional control in Escherichia coli
Acronym: Cas3 status
Leader: Ivana Ivančić Baće
Jurisdiction: Croatia
Funder: HRZZ
Funding stream: IP
Study programmeTitle: Graduate university studies in chemistry; specializations in: Research programme
Course: Research programme
Study programme type: university
Study level: graduate
Academic / professional title: magistar/magistra kemije (magistar/magistra kemije)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Embargo expiration date: 2022-07-15
Terms of useLicence In copyright icon
Created on2021-09-02 12:18:34