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Researches about Interaction and Genetic Transformation on Light Regulator of COP1, HY5

Author: SunMei
Tutor: LiYuHua
School: Northeast Forestry University
Course: Developmental Biology
Keywords: COP1 HY5 The light signal transduction Interaction Genetic Transformation
CLC: Q943
Type: Master's thesis
Year: 2009
Downloads: 196
Quote: 0
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Abstract


Light on the regulation of plant anthocyanin synthesis, mainly through the photoreceptors receive light signals for signal transduction. There are a lot of important signaling factors involved in light signal transduction. In this study, according to as test materials, light type varieties of anthocyanin synthesis Tsuda (Brassica rapa'Tsuda '), cloned COP1 and HY5 and the Arabidopsis light signal transduction factor highly similar gene, and its transcriptional expression features and the interaction between the two proteins provide a theoretical basis, in order to reveal the regulation of the synthesis of anthocyanins by light type light signal transduction mechanism. The following main results: 1 of Tsuda class BrCOP1, Cloning and Expression of BrHY5 by RACE and homologous PCR cloned from Tsuda similar to the Arabidopsis COP1, HY5 sequence genes BlastP analysis shows that with The Arabidopsis COP1 corresponding the HY5 sequence similarity over 90%, the domain projections indicate that the Ring and WD-40 domain containing COP1 and HY5 containing bZIP domain. Northern blot analysis showed that seedling, Tsuda, red, far-red light, blue, UV-B and UV-A treatment under COP1 and HY5 higher expression level was no significant difference in the amount of expression, which indicates that COP1 may HY5 participation in a the turnip seedlings above dregs Tin monochromatic light-induced reaction. COP1, HY5 in Tsuda mature plants no significant difference in the epidermis of leaves, flowers, taproot up to the amount of bait UV-A treatment taproot epidermis, COP1, HY5 expression increased. 2 Tsuda COP1 and HY5 interaction using GAL4 system, expression vectors, COP1 and HY5 were successfully constructed into an expression vector containing the binding domain and activation domain, after the sequencing of transformed yeast after defects medium, filtering and reporting genetic testing indicates that there is interaction between the turnip COP1 and HY5 protein. This result is consistent with the known results in Arabidopsis showed that Arabidopsis COP1 and HY5 similar functions optical signal transduction pathways may exist in the turnip. Expression vector 3 HY5-dsRNAi Construction and genetic study on the transformation to a more in-depth research turnip a HY5 the physiological functions, using the Gateway cloning system, primers based on cloned into the sediment Tin turnip HY5 gene sequence RNA interference was successfully constructed turnip the HY5 double-stranded RNA interference expression vector: HY5-dsRNAi. Agrobacterium-mediated transformation Tsuda, affecting turnip vitro regeneration of genetic transformation of various factors were studied. Optimization of the type of hormone factors, regeneration frequency of about 80% in vitro regeneration system to meet the requirements of genetic transformation. The study found that BA and NAA with more suitable Tsuda vitro regeneration, suitable for regeneration hormone combination of TDZ 7.0 mg / L NAA 1.0mg / L TDZ instead of their predecessors often used. 5.0 mg / LAgNO 3 turnip differentiation and regeneration is necessary.

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CLC: > Biological Sciences > Botany > Plant Cell Genetics
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