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A DNase footprinting assay [1] is a DNA footprinting technique from molecular biology/biochemistry that detects DNA-protein interaction using the fact that a protein bound to DNA will often protect that DNA from enzymatic cleavage. This makes it possible to locate a protein binding site on a particular DNA molecule.
DNase I Structure: DNase I is a glycoprotein with a molecular weight of 30,000 Da and a carbohydrate chain of 8-10 residues attached to Asn18 (orange). [3] It is an 𝛼,𝛽-protein with two 6-stranded 𝛽-pleated sheets which form the core of the structure. [4] These two core sheets run parallel, and all others run antiparallel.
In vivo footprinting is a technique used to analyze the protein-DNA interactions that are occurring in a cell at a given time point. [5] [9] DNase I can be used as a cleavage agent if the cellular membrane has been permeabilized. However the most common cleavage agent used is UV irradiation because it penetrates the cell membrane without ...
The study of DHS profiles combined with other techniques allows analysis of regulatory DNA in humans: Transcription factor: Using the ChIP-Seq technique, the binding sites to DNA in certain transcription factor groups are determined, and the DHS profiles are compared. The results confirm a high correlation, which show that the coordinated union ...
Deoxyribonuclease I (usually called DNase I), is an endonuclease of the DNase family coded by the human gene DNASE1. [5] DNase I is a nuclease that cleaves DNA preferentially at phosphodiester linkages adjacent to a pyrimidine nucleotide, yielding 5'-phosphate-terminated polynucleotides with a free hydroxyl group on position 3', on average producing tetranucleotides.
Diagnostic microbiology is the study of microbial identification. Since the discovery of the germ theory of disease , scientists have been finding ways to harvest specific organisms. Using methods such as differential media or genome sequencing , physicians and scientists can observe novel functions in organisms for more effective and accurate ...
Site-directed mutagenesis is a molecular biology method that is used to make specific and intentional mutating changes to the DNA sequence of a gene and any gene products. Also called site-specific mutagenesis or oligonucleotide-directed mutagenesis , it is used for investigating the structure and biological activity of DNA , RNA , and protein ...
Radioactive sulfur-35 was used to label the protein sections of the T2 phage, because sulfur is contained in protein but not DNA. [ 6 ] Hershey and Chase inserted the radioactive elements in the bacteriophages by adding the isotopes to separate media within which bacteria were allowed to grow for 4 hours before bacteriophage introduction.