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pTK-GLuc Vector |
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Description:
The pTK-Gluc Vector contains the coding sequence for the secreted Gaussia luciferase (GLuc) under the control of the Herpes Simplex Virus (HSV) thymidine kinase (TK) promoter, for constitutive expression in mammalian cells. It contains multiple cloning sites after the GLuc open reading frame (i.e. in the 3´ untranslated region) that can be used to insert heterologous sequences. Such constructs can be used for RNA studies including RNA stability, siRNA potency/efficacy, etc. by measuring the GLuc activity (1).
Gaussia luciferase is a new reporter luciferase from the marine copepod Gaussia princeps (2,3). This luciferase, which does not require ATP, catalyzes the oxidation of the substrate coelenterazine in a reaction that emits light (470 nm). It provides several advantages over other luminescent reporter genes:
• 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 (3).
• The secreted GLuc is very stable and can be stored for several days at 4°C without loss of activity.
Source:
Isolated from E. coli strain NEB 10-beta by a standard DNA purification procedure.
Advantages:- Constitutive expression of secreted GLuc
- MCS at the 3' untranslated region of GLuc
- Neomycin resistance
Concentration:
0.5 mg/ml
Storage Conditions
Storage Conditions:
10 mM Tris-HCl
1 mM EDTA
pH 7.5 @ 25°C
Storage Temperature:
-20°C
Restriction map of pTK-GLuc Vector. Only unique restriction sites are shown.
pTK-GLuc multiple cloning site (MCS). The Gaussia Luciferase sequence is shown with a blue background. Only unique restriction sites are shown.
References
- Morlighem, J.E., Petit, C. and Tzertzinis, G. (2007) Biotechniques, 42, 599-614.
- Verhaegen, 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.
Companion Products
Gaussia Luciferase Assay Kit
pCMV-GLuc Control Plasmid
pGLuc-Basic Vector
TransPass™ D2 Transfection Reagent
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