Difference between revisions of "Part:BBa J176013"

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VP64 is a transcriptional activator composed of four tandem copies of VP16 (Herpes Simplex Viral Protein 16, amino acids 437-447, DALDDFDLDML) connected with glycine-serine linkers. When fused to another protein domain that can bind near the promoter of a gene, VP64 acts as a strong transcriptional activator. This module is a classic molecular biology tool, which pre-dates fancy terms like "synthetic biology." See the [https://parts.igem.org/Part:BBa_J176013:Design References section] for some history on VP16 and VP64.
 
VP64 is a transcriptional activator composed of four tandem copies of VP16 (Herpes Simplex Viral Protein 16, amino acids 437-447, DALDDFDLDML) connected with glycine-serine linkers. When fused to another protein domain that can bind near the promoter of a gene, VP64 acts as a strong transcriptional activator. This module is a classic molecular biology tool, which pre-dates fancy terms like "synthetic biology." See the [https://parts.igem.org/Part:BBa_J176013:Design References section] for some history on VP16 and VP64.
  
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===Characterization===
 
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<b>VP64 activity is sensitive to promoter proximity.</b><br>
 
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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 (<partinfo>BBa_J176020</partinfo>) and targeted to Gal (UAS) DNA elements (<partinfo>BBa_J176019</partinfo>) that are assembled immediately adjacent to the HSVtkTATA minimal promoter (<partinfo>BBa_J176011</partinfo>), we observe weak activation, and perhaps repression, of a fluorescent reporter gene. In contrast, when a small spacer (<partinfo>BBa_J176039</partinfo>) is inserted between the Gal4 binding sites and the promoter, reporter gene activity greatly increases.
 
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(Data will be posted soon)
 
  
 
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Revision as of 02:48, 22 November 2011

VP64

Tetrameric VP16 transcription activator domain

Usage and Biology

  • Chassis: mammalian cells
  • Mammalian expression vector required
  • Protein domain; requires promoter, start codon, stop codon, and polyA signal for proper expression
  • VP64 doesn't do much as a free-floating domain. Fuse it to a DNA binding domain


VP64 is a transcriptional activator composed of four tandem copies of VP16 (Herpes Simplex Viral Protein 16, amino acids 437-447, DALDDFDLDML) connected with glycine-serine linkers. When fused to another protein domain that can bind near the promoter of a gene, VP64 acts as a strong transcriptional activator. This module is a classic molecular biology tool, which pre-dates fancy terms like "synthetic biology." See the References section for some history on VP16 and VP64.


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]