Expression, purification and binding of upstream regulatory proteins of GhCYC3

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dc.contributor Helsingin yliopisto, Maatalous-metsätieteellinen tiedekunta, Maataloustieteiden osasto fi
dc.contributor University of Helsinki, Faculty of Agriculture and Forestry, Department of Agricultural Sciences en
dc.contributor Helsingfors universitet, Agrikultur- och forstvetenskapliga fakulteten, Avdelningen för lantbruksvetenskaper sv
dc.contributor.author Das, Bishwajit
dc.date.issued 2020
dc.identifier.uri URN:NBN:fi:hulib-202012104915
dc.identifier.uri http://hdl.handle.net/10138/322786
dc.description.abstract Asteraceae comprises of approximately 10% of all angiosperm plant species. These species are well known for their highly compressed inflorescences known as capitula which consists of morphologically different types of flowers: ray, trans and disc flowers. This immense morphological difference excels Gerbera as an ideal plant to study flower type differentiations. Even though this complex process is governed by several genes, the ray flower identity is believed to be greatly influenced by GhCYC3 promoter mediated gene regulations. In previous studies two TCP transcription factors (TF): GhCIN1and GhCIN2, and two MADS TFs: GAGA1 and RCD5 were identified as the potential upstream regulators of GhCYC3. So, the aim of this study is to test whether these potential upstream regulators physically bind to GhCYC3 promoter in in vitro conditions. In order to achieve the goal, these transcription factor proteins from Gerbera hybrida were successfully expressed in E. coli and purified as fusion proteins to maltose-binding protein (MBP). Physical binding of the purified fusion proteins to the putative target DNA sites in the promoter region of GhCYC3 gene was tested by electrophoretic mobility shift assay (EMSA). The results showed that none of the gerbera transcription factors (GhCIN1, GhCIN2, GAGA1 and RCD5) bind to their putative target sites under the condition tested in this study. However, it might not be justifiable to deduce that these TFs do not interact with GhCYC3 promoter. The absence of in vitro interaction between the tested TFs and GhCYC3 promoter might be caused by either lack of proper folding and activity of the TFs or absence of co-factors which are available in vivo. en
dc.language.iso eng
dc.publisher Helsingin yliopisto fi
dc.publisher University of Helsinki en
dc.publisher Helsingfors universitet sv
dc.subject Asteraceae
dc.subject GhCYC3
dc.subject Transcription factor
dc.subject Protein purification
dc.subject TCP
dc.subject MADS
dc.title Expression, purification and binding of upstream regulatory proteins of GhCYC3 en
dc.type.ontasot pro gradu -tutkielmat fi
dc.type.ontasot master's thesis en
dc.type.ontasot pro gradu-avhandlingar sv
dc.subject.discipline Biotekniikka (MAAT) fi
dc.subject.discipline Biotechnology (MAAT) en
dc.subject.discipline Bioteknik (MAAT) sv
dct.identifier.urn URN:NBN:fi:hulib-202012104915

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