Immunology Technology Topics: Chromatin Immunoprecipitation Chips (Chip-Chip)
This technology enables rapid identification of specific binding sites for specific DNA-binding proteins in the chromosomes of the target genome. ChIP chips can also look for structural changes in chromosomes in any region of interest in a genome.
First, the use of ChIP-Chip
(1) Determining a DNA binding site of a gene transcription factor and a DNA binding site of another DNA binding protein or protein complex within a genome.
(2) Quantitative analysis of the chromosomal activity state.
(3) Functional study of histone modification. Changes in binding patterns associated with histone modifications can be determined by specific antibodies to acylated or methylated histones and specific antibodies to histones that have not been modified.
(4) Quantitative analysis of polymerase activity.
(5) Refine the biological information method and use the functional data to determine the position of the promoter.
Second, the specific experimental principle and experimental steps
Chromatin immunoprecipitation chip flow chart
Third, GeneChip-TilingArray technology introduction
On January 24, 2006, Affymetrix announced the launch of the GeneChip(R) human and mouse chimeric chip (TilingA, ray) series. This series of chips has been studied extensively beyond the known encoded protein sequences for systematic study of the entire human and mouse genome. Researchers can use this chip to study transcription factors and other protein binding domains. Recently, more researchers have used Affymetrix's chimeric chips to find many transcriptionally active regions that have never been found before in areas that were previously thought to be junk DNA. The TilingArray is by far the highest resolution gene chip type, and its probe design covers almost the entire sequence of the target DNA. To date, Affymetrix has developed a genome-wide Tiling chip for human, mouse, yeast, nematode, and Arabidopsis model organisms, providing a powerful analytical tool for studying the interaction of proteins and nucleic acids at the genome-wide scale. . In addition to the whole genome chip, GeneChip-TilingArray includes two human chip promoters and mouse promoters specifically used in ChIP-Chip technology. The probe design covers a range of 10kb near the transcription start site. The tumor-related 1 300 genes have increased their coverage to 12.5 kb.
In 1882, the German cytologist Fleming first published the observations of cell mitosis, and his work was regarded as one of the most important discoveries in the history of science. In addition to depicting and naming mitosis, Fleming also marks the chromatin, a substance that appears to play a key role in this process.
Currently, it is the focus of two of the most popular fields of biology - genomics and proteomics research. However, the difference is that Fleming uses an optical microscope and a glass bottle filled with a small amount of aniline to study it, and the latest genomic phase of chromatin research uses cutting-edge technology - chromatin immunoprecipitation Determination method (ChIP).
Both genomics and proteomics will use chromatin as a research object, but the methods used in the two fields vary. In genomics research, researchers usually start with a protein and use ChIP to find proteins associated with the genome. Proteomics studies use a reverse method in which a special DNA sequence is used as a bait to find proteins; then ChIP is used to confirm that those proteins are proteins that are associated with DNA sequences in vivo.
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