Calling cards for DNA-binding proteins

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Calling cards for DNA-binding proteins.

Identifying genomic targets of transcription factors is fundamental for understanding transcriptional regulatory networks. Current technology enables identification of all targets of a single transcription factor, but there is no realistic way to achieve the converse: identification of all proteins that bind to a promoter of interest. We have developed a method that promises to fill this void. ...

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"Calling cards" for DNA-binding proteins in mammalian cells.

The ability to chronicle transcription-factor binding events throughout the development of an organism would facilitate mapping of transcriptional networks that control cell-fate decisions. We describe a method for permanently recording protein-DNA interactions in mammalian cells. We endow transcription factors with the ability to deposit a transposon into the genome near to where they bind. Th...

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Calling Cards enable multiplexed identification of the genomic targets of DNA-binding proteins.

Transcription factors direct gene expression, so there is much interest in mapping their genome-wide binding locations. Current methods do not allow for the multiplexed analysis of TF binding, and this limits their throughput. We describe a novel method for determining the genomic target genes of multiple transcription factors simultaneously. DNA-binding proteins are endowed with the ability to...

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Topic Introduction Transposon Calling Cards

Identifying the genomic targets of transcription factors is an important step in understanding the regulatory networks of gene transcription in yeast. We have developed a method that utilizes what we refer to as transposon “calling cards,” in which a transcription factor directs the Ty5 retrotransposase to insert transposons into the genome adjacent to where the transcription factor binds. This...

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Gel Retardation Assays for DNA-binding Proteins.

MATERIALS 10x Tris-glycine buffer TBE buffer may be used in place of Tris-glycine. 6x Gel-loading buffer I 32P-labeled control DNA 32P-labeled target DNA of >20 bp (specific activity 2 x 107 cpm/μg [ Labeling of the DNA fragment can be accomplished by phosphorylation (Dephosphorylation of DNA Fragments with Alkaline Phosphataseor Phosphorylating the 5' Termini of Oligonucleotides), filling in o...

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ژورنال

عنوان ژورنال: Genome Research

سال: 2007

ISSN: 1088-9051

DOI: 10.1101/gr.6510207