Lecture12 - MCB 110L Spring 2010 Lecture 12 02/26/2010 Qing...

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Unformatted text preview: MCB 110L Spring 2010 Lecture 12 02/26/2010 Qing Zhong 306 Barker Hall How do we detect Cin8p in cells? Kerscher Lab, College of William and Mary Tag our favorite protein with green uorescent protein (GFP), which can be detected in vivo GFP expression in mammals GFP expression in Bacterialcell colonies GFP expression in yeast 2008 Nobel prize chemistry-GFP Osamu Shimomura , Martin Chalfie and Roger Y. Tsien Beta barrel protein scafold Fluorophore GFP a 238amino acid protein Osamu Shimomura, Marn Chale, Douglas Prasher, & Roger Y. Tsien GFP is rom the jellysh, A. victoria , and has a major excitaon peak at a wavelength o 395 nm and a minor one at 475 nm. Its emission peak is at 509 nm ( green visible spectrum ). Green Fluorescent Protein (GFP) Color of the emission spectrum is lower energy (longer wavelength) than the excitaOon spectrum. The chosen excitaOon wavelength determines the emission yield. Basic Fluorescence: ExcitaOon & Emission Spectra Shimomura first discovered GFP during the study of the bioluminescent protein aequorin, the mechanism by which certain jellyfish glow. In the footnote to his seminal paper on aequorin purification, he noted the additional presence of a protein giving solutions that look slightly greenish in sunlight through only yellowish under tungsten lights, and exhibiting a very bright, greenish fluorescence in the ultraviolet of a Mineralite, has also been isolated from squeezates. The frst guy- Martin Chalfie demonstrated the value of GFP as a luminous genetic tag for various biological phenomena. In one of his first experiments, he coloured six individual cells in the transparent roundworm Caenorhabditis elegans...
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This note was uploaded on 09/12/2011 for the course MCB 120L taught by Professor Fairclough during the Spring '08 term at UC Davis.

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Lecture12 - MCB 110L Spring 2010 Lecture 12 02/26/2010 Qing...

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