Chapter 6C 25 in addition to 27 February, 2004 Proteomics Structural in addition to Functional Charact
Peters, Brian, Meteorologist has reference to this Academic Journal, PHwiki organized this Journal Chapter 6C 25 in addition to 27 February, 2004 Proteomics Structural in addition to Functional Characterization in the Post-genomic era. Overview Proteomics examines the collection of proteins present in a given type of cell at a given time. Proteomics depends on genome sequences, but differs in the sense that the genome is static but the proteome changes over time. Global analysis of proteomes involves measuring a large number of phenotypes or concentrations simultaneously. The link between structure in addition to function is tight, but not well understood. Interactions between proteins as long as m networks of functionality. Defining these networks is the most challenging goal of proteomics.
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Pathway in addition to localization in as long as mation are available as long as the mTn mutant sets. Uptag/Downtag deletion strategy generates unique tags at deletion site. Unique tags are flanked by sites as long as PCR amplification.
Phenotype assignment using tagged deletion strategy Make a microarray containing UPTAG / DOWNTAG sequence. Each strain hybridizes to only one place on the microarray. Grow mixtures of hundreds of strains in the same flask under different conditions. Measure which strains grew by intensity of unique microarray hybridization of the PCR amplification of tags from all the strains present in the flask be as long as e in addition to after growth. Time zero = red, 6 hours later = green No difference -> Yellow, Defect = Red RNAi Small dsRNA complementary to a gene will make expression of the protein product of that gene impossible. Allows conditional knockouts in adult eukaryotes, especially C. elegans, where RNAi works even if the small dsRNA is eaten. RNAi is one of the most significant advances in functional genetics. New – first reported in 1998, just beginnning to be applied.
A high-throughput method to determine three dimensional crystal structure in addition to relate it to function is a long-term goal of proteomics: an example from the Archaea Aquaporin structure in addition to function Protein interactions Hard to detect since the rules are totally opaque Interaction depends on structure in complex ways. Empirical evidence is required. Coimmunoprecipitation High-throughput strategies Protein microarrays 2-hybrid interaction trap
See Math Minute 6.2 as long as a discussion of graph theory.
Which proteins are present Two-dimensional gel electrophoresis. Problems: Sample prep. Spot detection Quantification Identification
Assignment: Discovery Questions 1, 2, 5 ,6 ,9-13, 19, 20, 22-27, 35-39
Peters, Brian Meteorologist
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