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Thermo Scientific™ pTK-Gaussia Luc Vector for Luciferase Assays

Catalog No. PI16148
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PI16148 10 μg
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Catalog No. PI16148 Supplier Thermo Scientific™ Supplier No. 16148
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Description

Drive Gaussia luciferase expression as a reporter for gene regulation studies using vectors with a multiple cloning site (MCS) or control promoter (CMV or TK).

The Thermo Scientific™ pMCS-Gaussia Luc Vector is a multiple cloning site plasmid designed to accept a promoter sequence for study of gene regulation using the naturally secreting Gaussia luciferase reporter.

The Gaussia Luc Vectors contain a gene cloned from the copepod, Gaussia princeps. The gene encodes a naturally-secreted bioluminescent Gaussia luciferase (approx. 20kDa), which enables measurement of the reporter activity in media (for real-time assays) and in cell lysates. The pMCS vector contains a multiple cloning site for cloning a promoter to study its regulatory potential. The pCMV and pTK vectors have the luciferase gene under the CMV (Cytomegalovirus) promoter and Herpes Simplex Virus (HSV) thymidine kinase (TK) promoter, respectively. These constitutive expression vectors can be used as normalization controls to account for experimental variation in combination with other reporters.

Highlights:

  • Naturally-secreting Gaussia luciferase gene, optimized for high expression in mammalian systems
  • Multiple cloning site (MCS) provides versatility for transfer of regulatory elements from one plasmid to another
  • Transcription termination site (Ter), Lac operator (Lac O1), and transcriptional pause site (TPS) used to minimize background by reducing transcriptional read-through
  • Both puromycin (Pur) and ampicillin (Amp) markers for drug selection in mammalian and bacterial cells, respectively
  • High-copy pUC bacterial DNA replication origin
  • Two control vectors available with strong (CMV) or weak (TK) constitutive promoters for co-transfection and normalization

Specifications

Description pTK-Gaussia Luc Vector
Type Luc Vector
Cloning Method Restriction Enzyme/MCS
Delivery Type Transfection
Quantity 10 μg
Content And Storage Store in original container protected from direct sunlight in a dry, cool and well-ventilated area, between the following temperatures: -70 to -20°C.
Reporter Gene Gaussia Luciferase
Selection Agent (Eukaryotic) Puromycin
Product Line Pierce
For Use With (Application) Reporter Assays
Promoter TK
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Frequently Asked Questions (FAQs)

What options do you offer for luciferase vectors?

There are three vectors available for each expressed luciferase; pMCS, pCMV, and pTK. We recommend using pMCS to measure the activity of the reporter, and pCMV, and pTK as positive control vectors. For high level constitutive expression of Luciferase, we recommend using the CMV (Cytomegalovirus) promoter vector and for low level constitutive expression of luciferase, we recommend using the pTK (thymidine kinase) promoter vector.

What is the stability of the secreted luciferase?

Gaussia, Gaussia-Dura, and Cypridina Luciferase are stable in the culture media for greater than 24 hours. Generally, after transfection of CMV driven constructs, we see ever increasing secreted signal between 24 and 72 hours.
How quickly is luciferase secreted?

The earliest detectable signal will depend on the strength of the promoter. In general, with a strong promoter (such as CMV), significant signal over background can be seen in as little as 20 minutes. However, with a weaker inducible promoter, significant signal over background may take 1-2 hours after induction.
Do you still offer quantitative IP (qIP) Luciferase assay kits?

Our qIP Luciferase assay kits have been discontinued but we do carry epitope-tagged (HA or c-Myc) and Tluc (TurboLuc luciferase)-tagged mammalian expression vectors, and qIP protein interaction assay reagents for these assay kits.

For Research Use Only. Not for use in diagnostic procedures.