Transformacija uročnjaka Arabidopsis thaliana retrotranspozonom Tto1 primjenom bakterije Agrobacterium tumefaciens

master's thesis

• Title (english): Transformation of Arabidopsis thaliana with retrotransposon Tto1 by bacterium Agrobacterium tumefaciens
• Author: Igor Obleščuk
• Mentor: Zoran Zgaga (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Biochemical Engineering); (Laboratory for Biology and Microbial Genetics); Zagreb
• Defense date and country: 2006-12, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Istraživanja vezana uz aktivnost retrotranspozona u genomu domaćina često su ograničena zbog još nedovoljno poznatih mehanizama kojima stanice kontroliraju aktivnost tih elemenata. Zato se često retrotranspozone izolirane iz genoma jedne vrste nastoji ugraditi u genom druge. U svrhu istraživanja aktivnosti LTR-retrotranspozona duhana Tto1, konstruirana je modificirana kopija retrotranspozona Tto1. Promijenjenim retrotranspozonom transformirane su stanice uročnjaka Arabidopsis thaliana posredstvom bakterije Agrobacterium tumefaciens . Analizom je utvrđeno da je dio T-DNA u kojem se nalazio promijenjeni retrotranspozon Tto1 izgubljen delecijom u bakteriji A. tumefaciens prije transformacije stanica A. thaliana. Delecija je bila posljedica intramolekularne homologne rekombinacije između dva promotora Pnos i utvrđena je u svim stanicama. Uzrok neočekivano visokoj učestalosti utvrđene homologne rekombinacije u bakteriji A. tumefaciens nije otkriven.

Abstract (english)

Research of retrotransposon activity is often limited by insufficient knowledge of mechanisms by which the activity of these elements is controlled in the cells of the host genome. To circumvent this difficulty, retrotransposons from the host genome are often transferred to the genome of some other species. To investigate the activity of tobacco LTR-retrotransposon Tto1, in this work a modified version of Tto1 has been constructed. The modified Tto1 has been inserted into A. thaliana cells by agrobacterium-mediated transformation, followed by molecular analysis of transformants. This analysis revealed that the region of T-DNA harbouring the modified Tto1 was deleted with 100% eficency in A. tumefaciens cells, prior to transformation of A. thaliana. Although we established that the deletion is a consequence of intramolecular recombination, between the two Pnos promotors, the cause of extremely high recombination rate between these two promotors in A. tumefaciens remains unknown.

• Keywords: Agrobacterium tumefaciens; binarni vektor; intramolekularna homologna rekombinacija; retrotranspozon Tto1; transformacija
• Keywords (english): Agrobacterium tumefaciens; binary vector; intramolecular homologous recombination; retrotransposon Tto1; transformation
• Language: croatian
• URN:NBN: urn:nbn:hr:159:326365
• Study programme: Title: Biotechnology; specializations in: Biochemical-Microbiology; Course: Biochemical-Microbiology; Study programme type: university; Study level: graduate; Academic / professional title: diplomirani inženjer biotehnologije; smjer biokemijsko-mikrobiološki (diplomirani inženjer biotehnologije; smjer biokemijsko-mikrobiološki)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2017-09-08 09:08:09