Gaussia Luciferase (GLuc) is a recently discovered, naturally secreted protein by the copepod, Gaussia princeps (1). At 19kD, it is the smallest known luciferase and its catalytic properties make it one of the “brightest” known (2). Additionally, multiple disulfide bonds contribute to the protein stability at elevated temperatures. These properties make it an ideal reporter gene, either as a standalone expression monitor or as a fusion partner with other proteins (3). The first report of in vivo imaging using Gluc demonstrated sensitivity that is several orders of magnitude higher than the first generation luciferases, Firefly Luciferase (Fluc) and Renilla (4). In addition to mammalian cells, the secretion signal of GLuc is functional in all eukaryotic cells, including algae and nematodes. Scientists at NEB have used Gaussia luciferase to develop siRNA potency and miRNA expression assays (5,6). The GLuc reporter is increasingly being used to monitor the cellular secretory pathway and ER stress, to assay for viral infectivity and to study chemokine receptor activation (3,7-10).
- Balou, B., Szent-Gyorgyi, C. and Finley, G (2000) Abstract from 11th Int. Symp. on Biolumin. & Chemilumin. Asilomar, CA.
- Goerke, A., Loening, A., Gambhir, S. and Swartz, J. (2008) Metabolic Engineering, 10, 187–200. PMID: 18555198
- Hulleman JD et al.(2011) Mol. Biol. Cell December 15, 2011 vol. 22 no. 244765-4775. PMID: 22031286
- Tannous, B.A. et al (2005) Mol. Ther. 11, 435-443. PMID: 15727940
- Morlighem, J.E., Petit, C and Tzertzinis, G. (2007) Biotechniques, 42, 599-606. PMID: 17515198
- Shea C. and Tzertzinis G. unpublished observations (2007).
- Badr, et al. (2007) PloS ONE, 2, e571. PMID: 17593970
- Hewett, J.W., et al. (2007) Proc. Natl. Acad. Sci, USA, 104, 7271-7276. PMID: 17428918
- Capul, A. and de la Torre, J.C. (2008) Virology 382, 107-114. PMID: 18929379
- Luker, K., Gupta, M. and Luker G. (2009) Biotechniques, 47, 625-632. PMID: 19594447
Protocols for Gaussia Luciferase
- Luciferase Cell Lysis Buffer
- Protein expression using the K. lactis Protein Expression Kit - Cloning a PCR fragment into pKLAC2 (E1000).
- Protein Expression using the K. lactis Protein Expression Kit - Growth of strains for detection of secreted protein
- Protein Expression using the K. lactis Protein Expression Kit - Identification of Multi-copy Integrants
- Protein Expression using the K. lactis Protein Expression Kit - Identification of properly integrated cells
- Protein expression using the K. lactis Protein Expression Kit - Linearization of pKLAC2 for integrative transformation of K. lactis.
- Protein Expression using the K. lactis Protein Expression Kit - Simultaneous Expression of Multiple Proteins
- Protein Expression using the K. lactis Protein Expression Kit - Transformation of K. lactis GG799 cells
- Stabilized Assay Protocol I (Luminometers without injectors) (E3300)
- Stabilized Assay Protocol II (Injector-equipped luminometers) (E3300)
- Standard Assay Protocol I (Luminometers without injectors) (E3300)
- Standard Assay Protocol II (Injector-equipped luminometers) (E3300)
- Western Analysis (E8023)
- Naturally Secreted – Amenable to live cell assays
- Sensitivity – Brightest luciferases available; enables single cell applications
- Stability – Samples can be stored for several days with no loss of activity
- Easy-to-use – Cell lysis not necessary
- Non-destructive – Living cells can be used in downstream assays
- Flexible – Activity can also be measured in cell lysates
- Transfection optimization studies
- Promoter/enhancer assays
- High throughput assays
- Multiplex assays
- Multiple assays with other reporters
- Secretory pathway reporter assays
- Signal transduction
- siRNA potency screening
- Time course studies
- Single cell assays, including stem cells and primary cells
- Live cell assays
- Assays in difficult to transfect cells
NEB vs. Other Commercially Available Reporter Systems
|Does not require and
not affected by ATP
Comparison of the features of commercially available reporter systems highlights the advantages of using Gaussia or Cypridina Luciferase from NEB.