11-9-11_OmicsTraffickingA

11-9-11_OmicsTraffickingA - Cellular trafficking Kid #3:...

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Cellular trafficking Kid #3 : Hey, mister. Ain't you got a car? Eddie Valiant : Who needs a car in L.A.? We have the best public transportation system in the world. Judge Doom : A few weeks ago I had the good providence to stumble upon a plan of the city council. A construction plan of epic proportions. We're calling it a freeway. Eddie Valiant : Freeway? What the hell's a freeway? Judge Doom : Eight lanes of shimmering cement running from here to Pasadena. Smooth, safe, fast. Traffic jams will be a thing of the past. From “Who Framed Roger Rabbit?”
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Announcements Exam 4, Monday, November 14 at 8 pm in PHYS 114 and 112 Material from 10-25-11 (DNA repair) to today (Trafficking) will be covered on the exam.
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Recap Compare between conditions (disease, nutritional stress, development, etc.) Compare between species to determine their relationships Identify novel genes and proteins that are involved in a specific function How can we measure the expression of genes and proteins?
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DNA Sequencing This template DNA is supplied with a mixture of all four normal (deoxy) nucleotides in ample quantities dATP dGTP dCTP dTTP a mixture of all four dideoxy nucleotides, each present in limiting quantities and each labeled with a "tag" that fluoresces a different color: ddATP ddGTP ddCTP ddTTP DNA polymerase I Primer (DNA fragment complimentary to 3’ end of DNA) http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/D/DNAsequencing.html
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DNA sequence can be interpreted by a scanner - reads from bottom to top of the gel - can interpret the sequence according to the intensity and color of the detected fluorescence DNA Sequencing- Sanger-Coulson dideoxy method
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Construction of a gene-chip can measure changes in gene expression A collection of microscopic DNA spots, commonly representing single genes , are arrayed on a solid surface by covalent attachment to a chemical matrix. http://keck.med.yale.edu/affymetrix/genechip%20tile.jpg
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Dye labeled cDNA are added to the gene chip. cDNA that hybridize (bind) to the chip DNA cause that spot to glow Relative amount of red and green colors on a chip can be determined by a scanner. mRNA is first converted to more stable DNA, called cDNA, using reverse transcriptase
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Example: Which genes are expressed when in TCA cycle? http://shop.builder.hu/pictures/SAN_tca_cycle.jpg
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Does mRNA levels correlate with protein levels? Yes!
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This note was uploaded on 12/09/2011 for the course BIO 230 taught by Professor .. during the Fall '10 term at Purdue University-West Lafayette.

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11-9-11_OmicsTraffickingA - Cellular trafficking Kid #3:...

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