Protein domains/Reporters

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Introduction to Reporters

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...

Name	Description	AA sequence	Length
BBa_K165014	CFP x2		1494
BBa_K157004	Fluoresceine-A-binding		522
BBa_K3484002	Intein-mediated T3(THRB) → eGFP		2274
BBa_K3484000	Intein-mediated T3(THRB) → sfGFP		2268
BBa_K103009	linker fused to alpha fragment of TEM beta-lactamase		578
BBa_K103013	linker fused to omega fragment of TEM beta-lactamase		335
BBa_J176005	mCh		705
BBa_K165004	mCherry, yeast optimized for fusion		735
BBa_K103014	OmpA_linker_alpha_linker		1049
BBa_K103016	OmpA_linker_omega_linker		809
BBa_J70595	RFC12 Green Fluorescent Protein		711
BBa_K157024	SP-AntiNIP-GGGSlinker-Tm-betaLactamase(Bla1)		1488
BBa_K157025	SP-AntiNIP-GGGSlinker-Tm-betaLactamase(Bla2)		1233
BBa_K157028	SP-AntiNIP-GGGSlinker-Tm-Fluolinker-cCFP		1263
BBa_K157026	SP-AntiNIP-GGGSlinker-Tm-Fluolinker-cYFP		1263
BBa_K157029	SP-AntiNIP-GGGSlinker-Tm-nCFP		1428
BBa_K157027	SP-AntiNIP-GGGSlinker-Tm-nYFP		1431
BBa_K157031	SP-AntiNIP-GGGSlinker-Tm-Splitluciferase1/2		1239
BBa_K157030	SP-AntiNIP-GGGSlinker-Tm-Splitluciferase2/2		1239
BBa_K157032	SP-Lipocalin-GGGSlinker-Tm-betaLactamase(Bla1)		1275
BBa_K157033	SP-Lipocalin-GGGSlinker-Tm-betaLactamase(Bla2)		1020
BBa_K157036	SP-Lipocalin-GGGSlinker-Tm-Fluolinker-cCFP		1050
BBa_K157034	SP-Lipocalin-GGGSlinker-Tm-Fluolinker-cYFP		1050
BBa_K157037	SP-Lipocalin-GGGSlinker-Tm-nCFP		1215
BBa_K157035	SP-Lipocalin-GGGSlinker-Tm-nYFP		1218
BBa_K157039	SP-Lipocalin-GGGSlinker-Tm-Splitluciferase1/2		978
BBa_K157038	SP-Lipocalin-GGGSlinker-Tm-Splitluciferase2/2		1026
BBa_K5466030	Split mCerulean C-terminal. Saccharomyces cerevisiae codon optimized		273
BBa_K5466029	Split mCerulean N-terminal. Saccharomyces cerevisiae codon optimized		540
BBa_K157005	Split-Cerulean-cCFP		243
BBa_K157006	Split-Cerulean-nCFP		465
BBa_K157007	Split-Venus-cYFP		243
BBa_K157008	Split-Venus-nYFP		468
BBa_K157019	Tm-betaLactamase(Bla2)		363
BBa_K157018	Tm-betaLactamaseBla1		618
BBa_K157014	Tm-Fluo-linker-cYFP		393
BBa_K157016	Tm-Fluolinker-cCFP		393
BBa_K157017	Tm-nCFP		558
BBa_K157015	Tm-nYFP		561
BBa_K157020	Tm-Split-Luciferase1/2		321
BBa_K157021	Tm-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