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Slides to know (2) - Know the equations for resolution and...

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Unformatted text preview: Know the equations for resolution and Gibbs free energy (charged and uncharged molecules). e able to solve for any of the terms. Know these techniques, or what they are used for: PCR, cloning, gel electrophoresis (DNA and SDS), Southern blot, western blot, column chromatography, 2D gel, FRAP, single particle tracking, patch clamp Be able to solve for any of the terms. Know the chemical groups that each of the amino acids belong to (polar, nonpolar, etc) Know what the lipid groups look like Know the important amino acids in the K+ ion channel Light Microscopy Light diffracted through series of glass lenses to produce magnified image ight path: Light path: 1) Condenser lens 2) Specimen 3) Objective lens 4) Ocular lens 5) Eye or viewing screen Fig. 18.1 Maximum Magnification 1000 x light gathering ability (numerical aperture) of lens ow is the fluorescent probe attached to the specific protein? How is the fluorescent probe attached to the specific protein? 1) Express the specific protein in bacteria, purify it, and covalently link fluorescent molecule to it in vitro (in test tube) The labeled molecule is then injected into the cell viewed by epifluorescence microscopy (live or fixed cell) y p py ( ) 2) Immunofluorescence- Use a fluorescently labeled ntibody that recognizes the specific protein to stain antibody that recognizes the specific protein to stain a fixed specimen of the cell that is viewed by epifluorescence microscopy (fixed cell) 3) GFP-tagging of the protein- Express the protein as a GFP- fusion protein in a living cell that is viewed by epifluoresence p g y p microscopy (live or fixed cell) fluorescently labeled Method 2: Immunofluorescence anti-actin antibody Microscopy xed eukaryotic fixed eukaryotic cell fluorescently labeled Method 2: Immunofluorescence anti-actin antibody Microscopy xed eukaryotic fixed eukaryotic cell actin DNA coding sequence: GFP coding sequence: Method 3: GFP Tagging ATG GGG AAG GGA CCG TTG. ATG CCC CGT AAA ctin protein Actin protein GFP protein ctin FP sed coding sequence actin- GFP fused coding sequence ATG GGG AAG GGA CCG TTG CCC CGT AAA fusion gene inserted into genome of cell, rotein expressed p otei exp essed and localization observed in living cell Actin-GFP protein eye photographic film Uses beam of electrons Electron Microscopy instead of beam of light ectron n be reduced-electron can be reduced by accelerating it (Voltage applied)-electromagnet instead of glass lenses (Current applied)- pecimen coated with Specimen coated with heavy metals to get better diffraction of electrons Fig. 18.12 Maximum Mag =1000x na Maximum Mag = 250,000 x Maximum Res = 0.2 m Maximum Res = .001 m Level of Structure Definition Bonds Involved Comments Primary (1 ) Absolute sequence of amino acids from amino Peptide bonds All 3 higher levels are direct consequences of 1 structure (contains information about their final end to carboxyl end shape). Changes can lead to disease (ex. sickle cell) or little or no effect.little or no effect....
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This note was uploaded on 08/25/2009 for the course BIO 315 taught by Professor Steiner during the Spring '08 term at Kentucky.

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Slides to know (2) - Know the equations for resolution and...

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