Tagging of the chloroplast sensor kinase (CSK) with affinity epitopes HA and FLAG

Marinović, Mila (2011) Tagging of the chloroplast sensor kinase (CSK) with affinity epitopes HA and FLAG. Diploma thesis, Faculty of Science > Department of Biology.

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In plant genome of Arabidopsis thaliana (L.) Heynh gene At1g67840 that encodes chloroplast sensor kinase (CSK), which is a part of ancestral two-component signal systems has been identified. Sequence analysis showed high similarity of motifs in all major plant and algal lineages which indicates that CSK has evolved from a cyanobacterial sensor histidine kinase. The subject of this work has been tagging of the CSK protein with affinity epitopes HA and FLAG for the functional and structural characterization purpose of the sensor kinase and more specific deciphering of it’s role in Arabidopsis thaliana plant. For that purpose we fused epitopes HA and FLAG into the carboxy-terminal end of the CSK protein by polymerase chain reaction. Using “Gateway” recombinant technology CSK_HA_FLAG construct has been inserted into the entry and subsequently into the binary pH7WG2,0 vector. Futhermore, Agrobacterium EHA105 cells were transformed with a final construct pH7WG2,0-CSK_HA_FLAG with a goal of it further insertion in Arabidopsis thaliana plant genome, specifically into the CSK-KO plants.

Item Type: Thesis (Diploma thesis)
Keywords: Arabidopsis thaliana, CSK, two-component systems, sensor kinases, photosynthesis, “Gateway” technology
Supervisor: Fulgosi, Hrvoje
Date: 2011
Number of Pages: 59
Subjects: NATURAL SCIENCES > Biology
Divisions: Faculty of Science > Department of Biology
Depositing User: Grozdana Sirotic
Date Deposited: 18 Jul 2014 11:27
Last Modified: 18 Sep 2014 09:38
URI: http://digre.pmf.unizg.hr/id/eprint/2522

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