Generation of Arabidopsis thaliana (Heynh.) plants containing the TROL protein labeled with HA and FLAG protein markers

Kovačević, Jelena (2013) Generation of Arabidopsis thaliana (Heynh.) plants containing the TROL protein labeled with HA and FLAG protein markers. Diploma thesis, Faculty of Science > Department of Biology.

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Abstract

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.

Item Type: Thesis (Diploma thesis)
Keywords: floral dip, transformation, HA tag, photosynthesis, selection
Supervisor: Fulgosi, Hrvoje
Co-supervisor: Bauer, Nataša
Date: 2013
Number of Pages: 48
Subjects: NATURAL SCIENCES > Biology
Divisions: Faculty of Science > Department of Biology
Depositing User: Grozdana Sirotic
Date Deposited: 19 Feb 2014 13:06
Last Modified: 22 Sep 2014 11:43
URI: http://digre.pmf.unizg.hr/id/eprint/438

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