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master's thesis
Obilježavanje kloroplastne senzorske kinaze (CSK) afinitetnim biljezima HA i FLAG
Zagreb: University of Zagreb, Faculty of Science, 2011. urn:nbn:hr:217:509383
Marinović, Mila
University of Zagreb
Faculty of Science
Department of Biology

Institutional repository: Repository of the Faculty of Science

Cite this document

Marinović, M. (2011). Obilježavanje kloroplastne senzorske kinaze (CSK) afinitetnim biljezima HA i FLAG (Master's thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:509383

Marinović, Mila. "Obilježavanje kloroplastne senzorske kinaze (CSK) afinitetnim biljezima HA i FLAG." Master's thesis, University of Zagreb, Faculty of Science, 2011. https://urn.nsk.hr/urn:nbn:hr:217:509383

Marinović, Mila. "Obilježavanje kloroplastne senzorske kinaze (CSK) afinitetnim biljezima HA i FLAG." Master's thesis, University of Zagreb, Faculty of Science, 2011. https://urn.nsk.hr/urn:nbn:hr:217:509383

Marinović, M. (2011). 'Obilježavanje kloroplastne senzorske kinaze (CSK) afinitetnim biljezima HA i FLAG', Master's thesis, University of Zagreb, Faculty of Science, accessed 23 December 2024, https://urn.nsk.hr/urn:nbn:hr:217:509383

Marinović M. Obilježavanje kloroplastne senzorske kinaze (CSK) afinitetnim biljezima HA i FLAG [Master's thesis]. Zagreb: University of Zagreb, Faculty of Science; 2011 [cited 2024 December 23] Available at: https://urn.nsk.hr/urn:nbn:hr:217:509383

M. Marinović, "Obilježavanje kloroplastne senzorske kinaze (CSK) afinitetnim biljezima HA i FLAG", Master's thesis, University of Zagreb, Faculty of Science, Zagreb, 2011. Available at: https://urn.nsk.hr/urn:nbn:hr:217:509383

Metadata
TitleObilježavanje kloroplastne senzorske kinaze (CSK) afinitetnim biljezima HA i FLAG
Title (english)Tagging of the chloroplast sensor kinase (CSK) with affinity epitopes HA and FLAG
AuthorMila Marinović
MentorHrvoje Fulgosi (mentor)
Committee memberHrvoje Fulgosi (predsjednik povjerenstva)
Committee memberDunja Leljak-Levanić (član povjerenstva)
Committee memberZdravko Dolenec (član povjerenstva)
Committee memberBranka Pevalek-Kozlina (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Science
(Department of Biology)
Zagreb
Defense date and country2011, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Biology
Abstract
U genomu biljke Arabidopsis thaliana (L.) Heynh identificiran je gen At1g67840 koji kodira za protein kloroplastnu senzorsku kinazu (CSK), dio „drevnih“ dvokomponentnih signalnih sustava. Analizom aminokiselinskog slijeda utvrđena je visoka sačuvanost i sličnost određenih motiva u svih vaskularnih biljaka i algi te je protein CSK tip bakterijske senzorske histidinske kinaze utvrđene u fotosintetskim plastidima tj. kloroplastima. Cilj ovog rada je obilježavanje CSK proteina afinitetnim biljezima HA i FLAG u svrhu funkcionalne i strukturalne karakterizacije senzorske kinaze i specifičnijeg utvrđivanja njene uloge kod biljke Arabidopsis thaliana. Lančanom reakcijom polimerazom fuzionirali smo biljege HA i FLAG na karboksi terminalni kraj CSK proteina. Koristeći „Gateway“ rekombinacijsku tehnologiju konstrukt CSK_HA_FLAG ugrađuje se u ishodišni pENTR (engl. entry) vektor te naknadno u binarni pH7WG2,0 vektor. Nadalje, stanice agrobakterija EHA105 transformirane su s konstruktom pH7WG2,0-CSK_HA_FLAG s ciljem daljnje ugradnje CSK_HA_FLAG komponente prisutne u T-DNA regiji binarnog vektora u genom biljke Arabidopsis thaliana. Ovim postupkom u transformiranim biljkama CSK-KO (engl. knock-out), inaktiviran je gen At1g67840.
Abstract (english)
In plant genome of Arabidopsis thaliana (L.) Heynh gene At1g67840 that encodes chloroplast sensor kinase (CSK), which is a part of ancestral two-component signal systems has been identified. Sequence analysis showed high similarity of motifs in all major plant and algal lineages which indicates that CSK has evolved from a cyanobacterial sensor histidine kinase. The subject of this work has been tagging of the CSK protein with affinity epitopes HA and FLAG for the functional and structural characterization purpose of the sensor kinase and more specific deciphering of it’s role in Arabidopsis thaliana plant. For that purpose we fused epitopes HA and FLAG into the carboxy-terminal end of the CSK protein by polymerase chain reaction. Using “Gateway” recombinant technology CSK_HA_FLAG construct has been inserted into the entry and subsequently into the binary pH7WG2,0 vector. Futhermore, Agrobacterium EHA105 cells were transformed with a final construct pH7WG2,0-CSK_HA_FLAG with a goal of it further insertion in Arabidopsis thaliana plant genome, specifically into the CSK-KO plants.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:217:509383
Study programmeTitle: Biology; specializations in: Ecology
Course: Ecology
Study programme type: university
Study level: graduate
Academic / professional title: magistar/magistra biologije (magistar/magistra biologije)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Terms of useLicence In copyright icon
Created on2018-04-09 09:30:14