Difference between revisions of "Part:BBa J176013"

Revision as of 02:29, 16 October 2011

VP64

Tetrameric VP16 transcription activator domain

Usage and Biology

VP64 is a transcriptional activator composed of four tandem copies of VP16 (Viral Protein 16), which is derived from the herpes simplex virus. When fused to another protein domain that can interact directly with a gene (e.g., a DNA binding domain), VP64 acts as a strong transcriptional activator.

Characterization

VP64 activity is sensitive to promoter proximity.
In our hands, VP64 activity is maximized when it is targeted to DNA sequences that are ~40-80 base pairs away from a minimal promoter. When fused to the Gal4 DNA binding domain and targeted to Gal (UAS) DNA elements that are assembled immediately adjacent to the HSVTkTATA minimal promoter (BBa_J176011), we observe weak activation, and perhaps repression, of a fluorescent reporter gene. In contrast, when a small spacer is inserted between the Gal4 binding sites and the promoter, reporter gene activity greatly increases.

(Data will be posted soon)

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 9: / Line 9: @@
 ===Characterization===
 <b>VP64 activity is sensitive to promoter proximity.</b><br>
-In our hands, VP64 activity is diminished when it is targeted too close to a promoter. When fused to the Gal4 DNA binding domain and targeted to Gal (UAS) DNA elements that are assembled immediately adjacent to the HSVTkTATA minimal promoter ([part:BBa_J176011]), we observe weak activation, and perhaps repression, of a fluorescent reporter gene. In contrast, when a small spacer is inserted between the Gal4 binding sites and the promoter, reporter gene activity greatly increases.
+In our hands, VP64 activity is maximized when it is targeted to DNA sequences that are ~40-80 base pairs away from a minimal promoter. When fused to the Gal4 DNA binding domain and targeted to Gal (UAS) DNA elements that are assembled immediately adjacent to the HSVTkTATA minimal promoter (BBa_J176011), we observe weak activation, and perhaps repression, of a fluorescent reporter gene. In contrast, when a small spacer is inserted between the Gal4 binding sites and the promoter, reporter gene activity greatly increases.
 (Data will be posted soon)