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SRP2017

Sigma-Aldrich

GAL4 [(1-147) + VP16 (411-490)] from Saccharomyces cerevisiae human herpesvirus 2

recombinant, expressed in E. coli, ≥80% (SDS-PAGE)

Synonym(s):

VP16

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About This Item

UNSPSC Code:
12352200
NACRES:
NA.26

biological source

Saccharomyces cerevisiae
human herpesvirus 2

recombinant

expressed in E. coli

Assay

≥80% (SDS-PAGE)

form

frozen liquid

mol wt

~27.8 kDa

packaging

pkg of 10 and 500 μg

storage condition

avoid repeated freeze/thaw cycles

concentration

500 μg/mL

technique(s)

electrophoretic mobility shift assay: suitable

color

colorless to clear

NCBI accession no.

UniProt accession no.

shipped in

dry ice

storage temp.

−70°C

Gene Information

Saccharomyces cerevisiae ... GAL4(855828)
human herpesvirus 2 ... HS2VP16A(1487335)

General description

VP16 is a part of the tegument of herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) particles, and is a prototypical acidic activator. GAL4-VP16 is constructed by the fusion of the acidic activation domain of the HSV (herpes simplex virus) VP16 transactivator to the DNA binding domain of GAL4. VP16 (411-490) is the 89-residue activation domain of VP16 protein.

Biochem/physiol Actions

The GAL4 protein of yeast activates the transcription of several genes involved in galactose metabolism. This event requires that GAL4 bind to upstream activation sites with the consensus sequence 5′-CGGN5(T/A)N5CCG-3′ . A fragment of the GAL4 protein, comprising amino acids 1-147, binds DNA but fails to activate transcription. Herpes virus VP16 activates expression of immediate early genes in virally-infected cells. As most other eukaryotic transcriptional activator proteins, VP16 has a modular domain structure: it′s N-terminus is involved in DNA-protein interactions, while its C-terminal 79 amino acids have proven to be an especially potent transactivation domain. When fused to the DNA-binding domain of the yeast GAL4, this VP16 fragment functions as an activator of transcription in yeast, mammalian cells and in vitro transcription assays. VP16 has been shown to bind to TBP, TFIIB, and replication factor A.
The GAL4 yeast protein is responsible for the induction of various genes involved in galactose metabolism. This requires the binding of GAL4 gene to the upstream activation sites containing the consensus sequence 5′-CGGN5(T/A)N5CCG-3′. VP16 protein is essential for inducing the expression of viral immediate-early (IE) genes. A fragment of the GAL4 protein, comprising amino acids 1-147, binds DNA but fails to activate transcription. The highly acidic C-terminal tail of VP16 functions as a potent transcriptional activator in mammalian cells. GAL4-VP16 construct functions as an activator which promotes transcription in mammalian cells.

Physical form

Clear and colorless frozen liquid solution

Preparation Note

Use a manual defrost freezer and avoid repeated freeze-thaw cycles. While working, please keep sample on ice.

Storage Class Code

10 - Combustible liquids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Certificates of Analysis (COA)

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Large-scale chromatin unfolding and remodeling induced by VP16 acidic activation domain.
Tumbar T, et al.
The Journal of Cell Biology, 145(7), 1341-1354 (1999)
A vector for expressing GAL4(1-147) fusions in mammalian cells.
Sadowski I and Ptashne M
Nucleic Acids Research, 17(18) (1989)
A synergistic increase in potency of a multimerized VP16 transcriptional activation domain.
Emami KH1 and Carey M.
The Embo Journal, 11(13), 5005-5012 (1992)
Hydrophobic cluster analysis predicts an amino-terminal domain of varicella-zoster virus open reading frame 10 required for transcriptional activation.
Moriuchi H, et al.
Proceedings of the National Academy of Sciences of the USA, 92(20), 9333-9337 (1995)
L E Post et al.
Cell, 24(2), 555-565 (1981-05-01)
We report a system for investigating promoters of eucaryotic cell and virus genes based on analyses of the regulation of herpes simplex virus 1 (HSV-1) thymidine kinases whose structural gene sequences have been fused to the promoter of the gene

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