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pGLuc Mini-TK Vector |
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- Low Basal Expression
- MCS upstream of the secreted GLuc
- Extremely sensitive reporter
- Neomycin resistance
Description:
The pGLuc Mini-TK Vector contains a minimal promoter fragment from the HSV thymidine kinase promoter upstream of the Gaussia Luciferase coding sequence. The vector can be used for inserting transcriptional enhancer sequences in the MCS in order to assess their transcriptional activation functions. For transcriptional repression investigations the full HSV-TK promoter (pTK-GLuc) that gives higher basal activity should be used instead.
The pGLuc Mini-TK Vector contains the coding sequence of the secreted Gaussia Luciferase for expression in mammalian cells. Gaussia Luciferase is a new reporter luciferase from the marine copepod Gaussia princeps (1,2). This luciferase, which does not require ATP, catalyzes the oxidation of the substrate coelenterazine in a reaction that emits light (470 nm).
A DNA genomic fragment containing putative estrogen response elements was inserted into the polylinker of pGLuc Mini-TK. The properties of this fragment were assessed in a transfection assay. GLuc activity was measured in the culture supernatant of MCF-7 cells transfected with the construct and treated with vehicle control (C) or estradiol (E2).
Source:
Isolated from E. coli strain NEB 10-beta by a standard DNA purification procedure.
Advantages:- GLuc possesses a natural secretory signal and upon expression is secreted into the growth medium. Therefore, lysis of the cells is not necessary before assaying the activity. This allows for multiple samples to be assayed at different time points from the same transfected cells.
- GLuc generates over 1000-fold higher bioluminescent signal intensity than Firefly and Renilla Luciferases, making it an ideal sensitive transcriptional reporter. Additionally, it is much more sensitive than other secreted reporters such as SEAP or Metridia Luciferase (3).
- The secreted GLuc is very stable and can be stored for several days at 4°C without loss of activity.
Concentration:
500 μg/ml
Storage Conditions
Storage Conditions:
10 mM Tris-HCl
1 mM EDTA
pH 7.5 @ 25°C
Storage Temperature:
-20°C
Restriction map of pGLuc Mini-TK Vector. Only unique restriction sites are shown. The complete sequence and restriction map is available here.
In pGLuc Mini-TK Vector the MCS precedes the minimal TK promoter. The TATA box is underlined and the first amino acids of GLuc are indicated in the above sequence.
References
- Verhaegan, M. and Christopoulos, T.K. (2002) Anal. Chem., 74, 4378-4385.
- Tannous, B.A., Kim, D.E., Fernandez, J.L. Weissleder, R., and Breakfield, X.O. (2005) Mol. Ther., 11, 435-443.
- Wurdinger, X.X. et al. (2008) Nature Methods, 5, 171-173.
Companion Products
Gaussia Luciferase Assay Kit
pCMV-GLuc Control Plasmid
pGLuc-Basic Vector
pTK-GLuc Vector
TransPass™ D2 Transfection Reagent
Legal
Licenses/Patents/Disclaimers:
By opening this package or by using the materials enclosed within:
Recipient is legally bound and accepts the following terms and conditions:
Non-Commercial Entities: This product is covered by US Patent 6,232,107 and other patents that are legal property as assigned to Prolume Ltd./Nanolight Technologies. This product is licensed only to the purchasing laboratory-research group. Recipient agrees not to transfer this plasmid or derivatives of this vector to any other laboratory, person or research group, even if within the same institution.
Recipient agrees not to alter or make any changes to the nucleotide coding sequence or secretory coding sequence of the luciferase(s) contained within without prior written permission from Prolume Ltd./Nanolight Technologies (www.nanolight.com). Recipient agrees not to file for any patent rights or to any inventions claiming any portion of the luciferase(s) within this material without prior written permission from Prolume Ltd. Nanolight Technologies.
Commercial For-Profit Entities & Non-Profit Foundations (herein referred to as Commercial Recipients): Commercial Recipients wishing to derive products, engage in the sale or license of any products, discover drugs or make inventions by use of the materials enclosed, fully agree to the terms mentioned above for Non-Commercial Entities; AND ADDITIONALLY agree to and are hereby bound to use the materials FOR EVALUATION PURPOSES ONLY. Commercial Recipient hereby agrees to destroy and cease use of any materials or derivatives containing any portion of these materials within 120 days from receipt. Commercial Recipient agrees not to use the materials for any use, other than the 120 day suitability evaluation without prior written permission or obtaining a valid license from Prolume Ltd. Nanolight Technologies.
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