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Using BLAST to Analyze Proteins Using BLAST to Analyze Proteins

Using BLAST to Analyze Proteins - PowerPoint Presentation

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Using BLAST to Analyze Proteins - PPT Presentation

Part 2 Comparing Fluorescent Proteins 2013 Workshop C Cloning DNA to Make Proteins Dina N Kovarik MS PhD NWABR Fluorescent Proteins are Valuable Tools Locate proteins in the cell Track the migration of cells ID: 933599

proteins fluorescent protein source fluorescent proteins source protein gfp wikipedia http diversity red rfp related expressing sister mutations blue

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Slide1

Using BLAST to Analyze ProteinsPart 2: Comparing Fluorescent Proteins

2013 Workshop C: Cloning DNA to Make ProteinsDina N Kovarik, MS, PhDNWABR

Slide2

Fluorescent Proteins are Valuable Tools

Locate proteins in the cellTrack the migration of cellsReporter of expression

Sister

centromeres

(green)

mark the attachment of microtubules

(red)

to sister chromatids (blue). Left: Normal. Right: Drug-treated.

Source: http://mct.aacrjournals.org/content/2/5.cover-expansion

Mice expressing GFP under UV light (left & right), compared to normal mouse (center). Source: Wikipedia.

Slide3

Bioluminescence of the crystal jellyfish, Aequorea victoria

Source: http://www.conncoll.edu/ccacad/zimmer/GFP-ww/shimomura.html

238 amino acid proteins. GFP ribbon diagram. From PDB 1EMA.

Source: Wikipedia

Slide4

Rainbow of Fluorescent Proteins

Source: Wikipedia. http://en.wikipedia.org/wiki/File:FPbeachTsien.jpg

D

rawn

” with bacteria

expressing 8 different fluorescent proteins

Diversity of Mutations

 Diversity of Colors

mFruits

mBlueberry

(Blue Fluorescent Protein, or

BFP)

mLemon

(Yellow Fluorescent Protein, or

YFP)

mGrape1

(Cyan Fluorescent Protein, or

CFP)

Many

others, all with similarly ‘fruity’ names…

Slide5

Diversity and Key Mutations

Slide6

Research Questions

Is red fluorescent protein (RFP) related to its famous cousin, GFP, or is from a different source entirely? What other fluorescent proteins, if any, are closely related to RFP?

The cloning and protein purification experiments you have been conducting in the laboratory involve

mTomato

(related to

mCherry

), also called

red fluorescent protein (RFP).

Slide7