Biokemijska karakterizacija produkta gena At4g01150 iz biljke Arabidopsis thaliana

master's thesis

• Title (english): Biochemical characterization of At4g01150 gene product of Arabidopsis thaliana
• Author: Marta Čorić
• Mentor: Hrvoje Fulgosi (mentor)
• Mentor: Željka Vidaković-Cifrek (mentor)
• Committee member: Željka Vidaković-Cifrek (predsjednik povjerenstva)
• Committee member: Hrvoje Fulgosi (član povjerenstva)
• Committee member: Maja Matulić (član povjerenstva)
• Committee member: Nada Oršolić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2013-02-14, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Fotosinteza je značajan biološki proces u kojem se djelovanjem Sunčevog zračenja anorganske tvari (voda i ugljikov dioksid) prevode u složene organske spojeve i kisik, koji su bitni za prehranu skoro svih živih bića na Zemlji. Proteini uključeni u glavne reakcije fotosinteze najvećim su dijelom već otkriveni i njihove su funkcije istražene, no nedavnim sekvenciranjem genoma biljke Arabidopsis thaliana i brojnim sustavnim istraživanjima genoma biljaka te ekspresije proteina, pronađeni su novi... More proteini eksprimirani u biljnim kloroplastima, čije funkcije još nisu razjašnjene. Jedan od tih proteina je i produkt gena At4g01150 iz biljke A. thaliana, nazvan CAASE. Proučavanjem pojedinih odjeljaka kloroplasta pronađen je na vanjskoj i unutarnjoj membrani kloroplasta, u tilakoidima, te u plastoglobulima, no njegova funkcija do danas je nepoznata. Ovo istraživanje posvećeno je biokemijskim analizama proteina CAASE te dobivanju biljaka bez ekspresije CAASE-a. Protutijelo koje prepoznaje protein CAASE omogućilo je određivanje točne lokalizacije CAASE-a u biljnim tkivima koji proizvode fotosintezu, tj. sadrže kloroplaste, te u stroma tilakoidima kloroplasta. Biljke homozigoti za mutirani gen At4g01150 ipak proizvode male količine proteina CAASE, te se ne mogu koristiti u daljnjim istraživanjima. Zbog potreba izolacije i daljnjih istraživanja proteina CAASE konstruirana je rekombinantna molekula CAASE, s afinitetnim biljegom HA na C-terminalnom kraju proteina. Less

Abstract (english)

Oxygen photosynthesis is an important biological process in which the effect of solar energy can translate inorganic substances (water and carbon dioxide) into complex organic compounds, thus providing food and oxygen for all living organisms. Proteins involved in the main reactions of photosynthesis are mostly known, as well as their functions, but the recent Arabidopsis thaliana genome sequencing, and a number of systematic plant genome and protein expression studies, discovered new... More proteins localized in plant chloroplasts, whose functions are unclear. One of these proteins is the product of At4g01150 gene, named CAASE. The study of different compartments of chloroplasts showed that CAASE is present, on the outer and inner membranes of chloroplasts, thylakoids and in plastoglobules, but its function is so far unknown. In this study, the usage of the antibody that recognizes endogenous CAASE enabled the precise localization of the protein in photosynthetic plant tissues and, moreover, in the stroma thylakoids. However, the plants established to be devoid of CAASE still produced this protein, making it difficult to observe mutant phenotype. In order to investigate CAASE further, a vector for affinity purification was constructed. Less

• Keywords: kloroplasti; oksigena fotosinteza; genska inaktivacija; Gateway kloniranje; afinitetni proteinski biljezi; plava nativna elektroforeza
• Keywords (english): chloroplasts; oxygenic photosynthesis; gene inactivation; Gateway cloning; affinity protein tags; blue native electrophoresis
• Language: croatian
• URN:NBN: urn:nbn:hr:217:021510
• Study programme: Title: Molecular Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2017-04-25 12:51:30