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master's thesis
4
0
Generation of Arabidopsis thaliana (Heynh.) plants containing the TROL protein labeled with HA and FLAG protein markers
Jelena Kovačević (2013)
University of Zagreb
Faculty of Science
Department of Biology
Cite this item:
https://urn.nsk.hr/urn:nbn:hr:217:496612
Metadata
Title
Transgenične biljke Arabidopsis thaliana (Heynh.) koje eksprimiraju protein TROL obilježen proteinskim biljezima HA i FLAG
Author
Jelena Kovačević
Mentor(s)
Hrvoje Fulgosi
(thesis advisor)
Abstract
Transformacija biljaka metodom “floral dip” postupak je unosa T-DNA sekvence u genom stanica ovarija prije oplodnje. U ovom radu opisala sam transformaciju biljaka Arabidopsis thaliana molekulom T-DNA iz plazmidnog vektora pH7WG2,0 TROL-HA-FLAG. TROL je transmembranski protein koji sudjeluje u regulaciji puta prijenosa elektrona na tilakoidnim membranama kloroplasta. Dodatak proteinskih biljega HA i FLAG na protein TROL olakšava njegovu imunodetekcijsku provjeru i omogućuje daljnja istraživanja funkcije proteina TROL. Kako bih dokazala uspješnost transformacije biljaka genom za protein TROL-HA-FLAG, biljke sam analizirala metodama selekcije sjemenki na hranjivoj podlozi s antibiotikom, PCR provjere genoma i imunodetekcije proteina u listovima odraslih biljaka. U transformiranim biljkama sam imunodetekcijom potvrdila lokalizaciju proteina TROL-HA-FLAG u tilakoidnim membranama kloroplasta. Daljnja istraživanja omogućit će detaljniju potvrdu funkcionalnosti proteina TROL-HA-FLAG.
Keywords
floral dip
transformation
HA tag
photosynthesis
selection
Parallel title (English)
Generation of Arabidopsis thaliana (Heynh.) plants containing the TROL protein labeled with HA and FLAG protein markers
Committee Members
Nataša Bauer
(committee chairperson)
Hrvoje Fulgosi
(committee member)
Željka Vidaković-Cifrek
(committee member)
Dijana Škorić
(committee member)
Granter
University of Zagreb
Faculty of Science
Lower level organizational units
Department of Biology
Place
Zagreb
State
Croatia
Scientific field, discipline, subdiscipline
NATURAL SCIENCES
Biology
Study programme type
university
Study level
graduate
Study programme
Molecular Biology
Academic title abbreviation
mag. biol. mol.
Genre
master's thesis
Language
Croatian
Defense date
2013-02-05
Parallel abstract (English)
Floral dip is a transformation method that results in the insertion of T-DNA into the genome of unfertilized ovary cells. This thesis describes the transformation of Arabidopsis thaliana plants with T-DNA from pH7WG2,0 TROL-HA-FLAG plasmid vector. TROL is a transmembrane protein that regulates electron transport flow on the thylakoid membranes of chloroplasts. Addition of protein tags HA and FLAG to the TROL protein allows easier immunodetection analysis and enables further studies of the plant TROL protein function. In order to show the success of plant transformation with the TROL-HA-FLAG protein gene, the plants were subjected to multiple analyses such as seed selection method on growth plates with an added antibiotic, plant DNA PCR analysis and immunodetection analysis of plant leaf proteins. Due to the immunodetection analysis, I managed to confirm the localization of TROL-HA-FLAG protein in chloroplast thylakoid membranes of transformed plants. Further research could better show if TROL-HA-FLAG protein function is diminished.
Parallel keywords (Croatian)
„floral dip“
transformacija
HA epitop
fotosinteza
selekcija
Resource type
text
Access condition
Access restricted to students and staff of home institution
Terms of use
URN:NBN
https://urn.nsk.hr/urn:nbn:hr:217:496612
Committer
Palma Dizdarević