diplomsko delo
Alliana Kolar (Avtor), Ajda Taler-Verčič (Mentor)

Povzetek

Pomanjkanje joda v človekovem organizmu povzroča številne zdravstvene težave. Med njimi je najpogostejša golša; pomanjkanje hkrati povezujemo tudi z motnjami v razvoju otroka in možnostjo spontanega splava med nosečnostjo. Najvišje količine joda se nahajajo v morski hrani, v naših kuhinjah pa ga največ najdemo v namizni soli. Kljub temu pa ga večina ljudi ne zaužije v zadostni količini. Rešitev za to bi lahko bila biofortifikacija, kjer bi v rastlinske pridelke vnesli dovolj visoke količine joda. Ker pa kopenskim rastlinam jod ne predstavlja tako pomembne vloge v njihovem organizmu, bi to lahko bil problem, ki ga je treba preštudirati. Encima PorA in CYP18-3 najdemo v rastlinah in oba igrata pomembno vlogo pri njenem razvoju – oba povezuje pomembna vloga v procesu fotomorfogeneze. V rastlini ju najdemo tudi v jodirani obliki (jod se veže na tirozine), ni pa znano, ali ima jodiranje kakšen vpliv na njuno funkcijo. Da bi ugotovili, ali se aktivnost obeh encimov spremeni po jodiranju, smo ju želeli pripraviti kot rekombinantna proteina. Z molekulskim kloniranjem smo uspeli pripraviti le zapis za protein CYP18-3 in ga vstaviti v vektor pET-32a(+). Zapisu smo dodali N-končno heksahistidinsko oznako ter prepoznavno mesto za proteazo TEV. Ob tem smo uporabili bakterijski klonirni sistem E. coli DH5α. Proteina nato nismo uspeli pripraviti v izbranem bakterijskem ekspresijskem sistemu E. coli BL21[DE3] pLysS. Iz naših rezultatov ni razvidno ali sploh prode do ekspresije proteina, ali pa pride do razgradnje. Postopek bo potrebno še optimizirati.

Ključne besede

jodiranje;rastlinski proteini;PorA;CYP18-3;molekulsko kloniranje;biofortifikacija;diplomska dela;

Podatki

Jezik: Slovenski jezik
Leto izida:
Tipologija: 2.11 - Diplomsko delo
Organizacija: UL FKKT - Fakulteta za kemijo in kemijsko tehnologijo
Založnik: [A. Kolar]
UDK: 577.15:546.15(043.2)
COBISS: 171494659 Povezava se bo odprla v novem oknu
Št. ogledov: 74
Št. prenosov: 8
Ocena: 0 (0 glasov)
Metapodatki: JSON JSON-RDF JSON-LD TURTLE N-TRIPLES XML RDFA MICRODATA DC-XML DC-RDF RDF

Ostali podatki

Sekundarni jezik: Angleški jezik
Sekundarni naslov: Expression and isolation of plant enzymes PorA and CYP18-3
Sekundarni povzetek: Iodine deficiency is one of the most common deficiencies in the human body, causing various health problems. Among them, the most common is goiter; but it also causes disturbances in the development of a child and a possibility of spontaneous abortion during pregnancy. The highest amounts of iodine are found in seafood, but in our kitchens it is mostly found in table salt. However, most people do not consume enough of it. The solution for this problem could be biofortification, where sufficiently high amounts of iodine would be introduced into plant products. However, iodine does not represent such an important role for land plants in their organism, which could be a problem that needs to be researched. PorA and CYP18-3 enzymes are found in plants and both play an important role in its development – both are linked to an important role in the photomorphogenesis process. Both proteins are also found in the plant in iodinated form (iodine binds to tyrosines), but it is not known if iodination has any effect on their function. In order to determine whether the activity of both enzymes changes after iodination, we wanted to prepare it as a recombinant protein. By molecular cloning, we managed to prepare only a transcript for the CYP18-3 protein and insert it into the pET-32a(+) vector. An N-terminal hexahistidine tag and a recognition site for the TEV protease were added to the transcript. We used the bacterial cloning system E. coli DH5α. We were not successful to prepare the protein in the selected bacterial expression system E. coli BL21[DE3] pLysS. It is not clear from our results whether protein expression occurs at all, or whether degradation occurs. The process will still need to be optimized.
Sekundarne ključne besede: iodination;plant proteins;PorA;CYP18-3;molecular cloning;Encimi;Jod;Univerzitetna in visokošolska dela;
Vrsta dela (COBISS): Diplomsko delo/naloga
Študijski program: 1000371
Konec prepovedi (OpenAIRE): 1970-01-01
Komentar na gradivo: Univ. v Ljubljani, Fak. za kemijo in kemijsko tehnologijo, UNI Biokemija
Strani: 37 str.
ID: 19912970