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Latest revision as of 00:06, 9 December 2009

< Back to Protein domains

Introduction to Reporters

Cambridge 2009-RAINBOW.png

What are reporters? Why call them reporters and not fluorescent proteins, beta-galactosidase/Xgal assays or pigments? The answer is that in synthetic biology, they are often used as a mechanism to display information. If you design a biosensor system that is sensitive to substrate A, you ideally want a way of displaying these results without needing an additional assay. Using a reporter protein is a great way of achieving this property.

Reporter protein coding sequences encode proteins whose presence in the cell or organism is readily observed. For example, fluorescent proteins cause a cell to fluoresce when excited with light of a particular wavelength, luciferases cause a cell to catalyze a reaction that produces light, and enzymes such as beta-galactosidase convert a substrate to a colored product. Reporters are frequently used to quantify the strength or activity of upstream gene expression parts such as promoters and ribosome binding sites. Reporters, when fused in frame to other protein coding sequences, can also be used to identify where a protein is located in a cell or organism.

There are several different ways to measure or quantify a reporter depending on the particular reporter and what kind of characterization data is desired. Generally speaking, microscopy is useful for obtaining both spatial and temporal information on reporter activity, particularly at the single cell level. Flow cytometers are best suited for measuring the distribution in reporter activity across a large population of cells. Plate readers are generally best for taking population average measurements of many different samples over time.

Some iGEM teams have excellent projects that create new reporters, or make great use of existing ones. Please see the tables below for fluorescent proteins, chromoproteins, luciferase proteins and enzymes that produce colored substrates in the Registry.


More...
NameDescriptionAA sequenceLength
BBa_K165014CFP x2 1494
BBa_K157004Fluoresceine-A-binding  522
BBa_K3484002Intein-mediated T3(THRB) → eGFP  2274
BBa_K3484000Intein-mediated T3(THRB) → sfGFP 2268
BBa_K103009linker fused to alpha fragment of TEM beta-lactamase 578
BBa_K103013linker fused to omega fragment of TEM beta-lactamase 335
BBa_J176005mCh 705
BBa_K165004mCherry, yeast optimized for fusion 735
BBa_K103014OmpA_linker_alpha_linker 1049
BBa_K103016OmpA_linker_omega_linker 809
BBa_J70595RFC12 Green Fluorescent Protein 711
BBa_K157024SP-AntiNIP-GGGSlinker-Tm-betaLactamase(Bla1) 1488
BBa_K157025SP-AntiNIP-GGGSlinker-Tm-betaLactamase(Bla2) 1233
BBa_K157028SP-AntiNIP-GGGSlinker-Tm-Fluolinker-cCFP 1263
BBa_K157026SP-AntiNIP-GGGSlinker-Tm-Fluolinker-cYFP  1263
BBa_K157029SP-AntiNIP-GGGSlinker-Tm-nCFP 1428
BBa_K157027SP-AntiNIP-GGGSlinker-Tm-nYFP  1431
BBa_K157031SP-AntiNIP-GGGSlinker-Tm-Splitluciferase1/2  1239
BBa_K157030SP-AntiNIP-GGGSlinker-Tm-Splitluciferase2/2  1239
BBa_K157032SP-Lipocalin-GGGSlinker-Tm-betaLactamase(Bla1) 1275
BBa_K157033SP-Lipocalin-GGGSlinker-Tm-betaLactamase(Bla2) 1020
BBa_K157036SP-Lipocalin-GGGSlinker-Tm-Fluolinker-cCFP  1050
BBa_K157034SP-Lipocalin-GGGSlinker-Tm-Fluolinker-cYFP  1050
BBa_K157037SP-Lipocalin-GGGSlinker-Tm-nCFP  1215
BBa_K157035SP-Lipocalin-GGGSlinker-Tm-nYFP  1218
BBa_K157039SP-Lipocalin-GGGSlinker-Tm-Splitluciferase1/2  978
BBa_K157038SP-Lipocalin-GGGSlinker-Tm-Splitluciferase2/2  1026
BBa_K5466030Split mCerulean C-terminal. Saccharomyces cerevisiae codon optimized 273
BBa_K5466029Split mCerulean N-terminal. Saccharomyces cerevisiae codon optimized 540
BBa_K157005Split-Cerulean-cCFP 243
BBa_K157006Split-Cerulean-nCFP 465
BBa_K157007Split-Venus-cYFP 243
BBa_K157008Split-Venus-nYFP 468
BBa_K157019Tm-betaLactamase(Bla2) 363
BBa_K157018Tm-betaLactamaseBla1 618
BBa_K157014Tm-Fluo-linker-cYFP 393
BBa_K157016Tm-Fluolinker-cCFP 393
BBa_K157017Tm-nCFP 558
BBa_K157015Tm-nYFP 561
BBa_K157020Tm-Split-Luciferase1/2 321
BBa_K157021Tm-Splitluciferase2/2 369


References

<biblio>

  1. Pardy pmid=7866866

</biblio>

Fluorescent proteins

<biblio>

  1. Chalfie pmid=8303295
  2. Prasher pmid=1347277
  3. Cormack pmid=8707053
  4. Tsien pmid=9759496
  5. Andersen pmid=9603842
  6. Leveau pmid=11698362
  7. Iafolla pmid=18350571
  8. Dong pmid=18352052
  9. Bagh pmid=18352063
  10. Baird pmid=11050229
  11. Patterson pmid=15583657

</biblio>

Luciferase

<biblio>

  1. Bronstein pmid=8080073

</biblio>

β-galactosidase

<biblio>

  1. Griffith pmid=11779182
  2. Villarejo pmid=4552692
  3. Zamenhof pmid=4552986
  4. Ullman pmid=1345751

</biblio> [http://openwetware.org/index.php?title=Beta-Galactosidase_Assay_%28A_better_Miller%29 Miller Assay on OpenWetWare] by Sean Moore