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Microplate-nucleotide Hybridization Based Elisa-like Assay for Detection of DNA Binding Protein

Author: ZhangWenJuan
Tutor: ZengLingWen
School: University of Science and Technology of China
Course: Biochemistry and Molecular Biology
Keywords: DNA-binding proteins Transcription factor High stringency Microplate nucleic acid hybridization Enzyme - linked immunosorbent assay
CLC: Q503
Type: Master's thesis
Year: 2011
Downloads: 23
Quote: 0
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Abstract


Transcription factors , also known as the sequence-specific DNA binding proteins, containing one or more DNA-binding domains ( DBDS ) . They are localized in the cytoplasm , in state normal cell function or by a specific factor-induced , from the cytoplasm into the nucleus , with the cis-acting elements in the genome , such as promoter , enhancer , attenuator , specifically recognized constituent genes transcription machinery , thus achieving its function to regulate gene expression . Detection of expression and the degree of activation of the transcription factor , therefore, has a crucial role to our understanding of cell function . The detection technology is primarily based on the level of DNA and protein-protein interactions , namely the use of DNA as a probe to analyze the DNA binding activity of the transcription factor containing a cis-acting elements . The subject \the detection is converted to the detection of the specific nucleic acid sequence corresponding transcription factor . The method does not require complex instrumentation , tedious steps and specific antibodies to quantitatively study the degree of expression and activation of the transcription factor , and can be completed in a relatively short period of time the detection of the transcription factor . Detecting transcription factors c-Jun activity as a model to evaluate the detection method , in a relatively short period of time , the method can accurately detect of 3.5ng the transcription factor c - Jun pure protein and 0.625ug Hela nuclei crude extract, and has high sensitivity . Moreover , the detection method can be used for the detection of any of a transcription factor . Given based microplate nucleic acid hybridization, enzyme-linked immunosorbent assay \scientific research , such as cellular and molecular biology on a variety of processing the stimulus response screening aspects of evolutionary biology research , environmental monitoring and drug screening , and so on .

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CLC: > Biological Sciences > Biochemistry > General issues > Biochemical techniques
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