Difference between revisions of "Part:BBa K607003"
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GFP with LVA-tag under transcriptional control of a modified Prm(PcI) promoter.
 
GFP with LVA-tag under transcriptional control of a modified Prm(PcI) promoter.
 
The Prm promotor is induced by cI. For more information on cI, see: https://parts.igem.org/wiki/index.php?title=Part:BBa_K607001<br>
 
The Prm promotor is induced by cI. For more information on cI, see: https://parts.igem.org/wiki/index.php?title=Part:BBa_K607001<br>
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The Prm promotor is part of an operator system where cI or cro can be produced.
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Activation of PRM involves an interaction between an acidic patch on the surface of the helix–turn–helix motif of cI protein and a target region near the C terminus of the σ70 subunit of Escherichia coli RNA polymerase.
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<br>[[Image:operatorcI.jpg]]
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The GFP protein contains an LVA tag, so it will be degraded faster by proteolytic enzymes compared to GFP proteins without LVA tag.
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<br>cI dimers will bind to the first two operators. This will look like this:
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<br>
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<br>[[Image:2operatorcI.jpg]]
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<br>References
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<br>The “free” PRM promoter will subsequently be occupied by RNA polymerase which in turn will transcribe cI. This leads to a continuous transcription (DNA to RNA) and subsequent expression (RNA to protein) of cI. This means cI is regulating its own synthesis and simulataneously maintaining the repression of cro. Conversely this auto-regulation can also be obeserved if cro is abundant: cro dimer binding to the operator sites will block binding of RNA polymerase to PRM, but will allow binding to PR, which in turn will lead exclusively to cro transcription. At increasing concentrations of cI the probability of cI binding to OR3 also increases, inhibiting therefore RNA polymerase binding to PRM. This means at higher concentrations cI is inhibiting its own synthesis, acting as a self-repressor.
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<br> Changing the mechanism of transcriptional activation by phage λrepressor. Mei Li,* W. R. McClure,† and Miriam M. Susskind. Proc Natl Acad Sci U S A. 1997.  
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Revision as of 19:18, 21 September 2011

Prm_GFP-LVA

GFP with LVA-tag under transcriptional control of a modified Prm(PcI) promoter. The Prm promotor is induced by cI. For more information on cI, see: https://parts.igem.org/wiki/index.php?title=Part:BBa_K607001
Activation of PRM involves an interaction between an acidic patch on the surface of the helix–turn–helix motif of cI protein and a target region near the C terminus of the σ70 subunit of Escherichia coli RNA polymerase. The GFP protein contains an LVA tag, so it will be degraded faster by proteolytic enzymes compared to GFP proteins without LVA tag.

References
Changing the mechanism of transcriptional activation by phage λrepressor. Mei Li,* W. R. McClure,† and Miriam M. Susskind. Proc Natl Acad Sci U S A. 1997.

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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 752