Expression, purification and binding of upstream regulatory proteins of GhCYC3

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http://urn.fi/URN:NBN:fi:hulib-202012104915
Titel: Expression, purification and binding of upstream regulatory proteins of GhCYC3
Författare: Das, Bishwajit
Medarbetare: Helsingfors universitet, Agrikultur- och forstvetenskapliga fakulteten, Avdelningen för lantbruksvetenskaper
Utgivare: Helsingin yliopisto
Datum: 2020
Språk: eng
Permanenta länken (URI): http://urn.fi/URN:NBN:fi:hulib-202012104915
http://hdl.handle.net/10138/322786
Nivå: pro gradu-avhandlingar
Ämne: Biotekniikka (MAAT)
Biotechnology (MAAT)
Bioteknik (MAAT)
Abstrakt: 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.
Subject: Asteraceae
GhCYC3
Transcription factor
Protein purification
TCP
MADS


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