Kloniranje i uvođenje gena CgKCS za elongazu nezasićenih masnih kiselina iz grčke režuhe (Cardamine graeca L.) u stanice kvasca i bundeve

master's thesis

• Title (english): Cloning and expression of the gene CgKCS for fatty acid elongase from Cardmine graeca L. in yeast and pumpkin cells
• Author: Marijana Vrdoljak
• Mentor: Jasna Vorkapić-Furač (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Biochemical Engineering); (Laboratory for Cell Technology, Application and Biotransformations); Zagreb
• Defense date and country: 2011-07-08, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Nervonska kiselina je dugolančana mononezasićena masna kiselina prisutna u sfingolipidnoj frakciji bijele tvari mozga i u mijelinskom sloju perifernih živčanih vlakana sisavaca. Prirodan izvor te kiseline su rijetke biljne vrste. Zbog mnogobrojnih pozitivnih učinaka na opću kliničku sliku osoba s raznim neurološkim oboljenjima, kao i prerano rođene djece koja se ne hrane majčinim mlijekom, nervonska kiselina kao potencijalni bioaktivni lipidni dodatak prehrani jedan je od najperspektivnijih nutraceutika. Cilj ovoga rada bio je klonirati gen CgKCS koji kodira enzim elongazu dugolančanih masnih kiselina iz grčke režuhe (Cardamine graeca L.). Funkcionalnost genskog konstrukta za elongazu masnih kiselina provjerena je induciranom ekspresijom u transformiranim stanicama kvasca (Saccharomyces cerevisiae), a metodom plinske kromatografije dokazane su razlike u profilu masnih kiselina i prisustnost nervonske kiseline u transformiranom soju u odnosu na divlji soj kvasca. Cilj daljnjih istraživanja je, primjenom dobivenih konstrukata, postići gensku transformaciju bundeve, te na taj način obogatiti ulje bundeve nervonskom kiselinom.

Abstract (english)

Nervonic acid is a very long-chain monounsaturated fatty acid widely distributed in brains and nerve fibers of vertebrates. Natural source of nervonic acids are seeds of only a few plant speciesm Due to many positive effects on persons with multiple sclerosis and prematurely born formula-fed infants, nervonic acid is proposed to be evaluated as nutriceutic for human beings and animals. Aim of this study was to clone gene for a fatty acid elongase from Cardamine graeca. Functional analysis of CgKCS gene was confirmed by its expression in yeast (Saccharomyces cerevisiae). Gas chromatography analyses of fatty acids in yeast cells upon KCS expression revealed the presence of saturated and monounsaturated very long-chain fatty acids, including nervonic acid, that are not normally present in yeast. Further aim is to achieve genetic transformation of pumpkin by using the constructs obtained in this work, and to engineer nervonic acid-rich oil in the seed of pumpkin.

• Keywords: Cardamine graeca L.; Cucurbita pepo L. var Styriaca; gen KCS; nervonska kiselina; Saccharomyces cerevisiae
• Keywords (english): Cardamine graeca L.; Cucurbita pepo L. var Styriaca; gene KCS; nervonic acid; Saccharomyces cerevisiae
• Language: croatian
• URN:NBN: urn:nbn:hr:159:377574
• Study programme: Title: Molecular Biotechnology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra inženjer/inženjerka molekularne biotehnologije (magistar/magistra inženjer/inženjerka molekularne biotehnologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2017-05-31 12:22:02