Difference between revisions of "Part:BBa J176013:Experience"
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'''VP64 activity is sensitive to promoter proximity.'''<br>
 
'''VP64 activity is sensitive to promoter proximity.'''<br>
 
This is an "oops" discovery that I first learned about from Tom Armel and Jason Lohmeuller (P.A. Silver lab), then reproduced through improving one of my failed inducible promoters. In my 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>), I observed 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. (Data will be posted soon)
 
This is an "oops" discovery that I first learned about from Tom Armel and Jason Lohmeuller (P.A. Silver lab), then reproduced through improving one of my failed inducible promoters. In my 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>), I observed 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. (Data will be posted soon)
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Revision as of 21:03, 18 February 2012

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_J176013

User Reviews

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Karmella Haynes

VP64 activity is sensitive to promoter proximity.
This is an "oops" discovery that I first learned about from Tom Armel and Jason Lohmeuller (P.A. Silver lab), then reproduced through improving one of my failed inducible promoters. In my 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 (BBa_J176020) and targeted to Gal (UAS) DNA elements (BBa_J176019) that are assembled immediately adjacent to the HSVtkTATA minimal promoter (BBa_J176011), I observed weak activation, and perhaps repression, of a fluorescent reporter gene. In contrast, when a small spacer (BBa_J176039) is inserted between the Gal4 binding sites and the promoter, reporter gene activity greatly increases. (Data will be posted soon)


UNIQ71e0d718ff024b32-partinfo-00000006-QINU