Difference between revisions of "Part:BBa K227007:Design"

 
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The first part of our characterization begins with the puc promoter, which promotes transcription of the LH2 pucB/A genes naturally in ''Rhodobacter sphaeroides''.  The absorption spectra of a DBCOmega mutant (LH2 deficient) transformed with pRKCBC3 containing the puc promoter and pucB/A genes will allow us to characterize the puc promoter under high and low oxygen conditions. 
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More absorption of light at the LH2 spectra peaks normalized to culture OD corresponds with more transcription and vis versa.
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'''Method'''<br>
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The optical density of the cultures was recorded at 600nm, and each absorbance spectrum was normalized to OD600 by division by this value.  Background subtraction of spectrophotometer data was performed in Origin 6.1 Software.  A ten-point baseline was created by a "positive peak" algorithm then modified to approximate the scattering curve that falls as the inverse fourth power of wavelength.  <br>
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'''Results'''<br>
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[[Image:02 puc pro characterization graph.png]]
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<partinfo>BBa_K227007 short</partinfo>
 
<partinfo>BBa_K227007 short</partinfo>
  

Revision as of 22:27, 18 October 2009


The first part of our characterization begins with the puc promoter, which promotes transcription of the LH2 pucB/A genes naturally in Rhodobacter sphaeroides. The absorption spectra of a DBCOmega mutant (LH2 deficient) transformed with pRKCBC3 containing the puc promoter and pucB/A genes will allow us to characterize the puc promoter under high and low oxygen conditions. More absorption of light at the LH2 spectra peaks normalized to culture OD corresponds with more transcription and vis versa.

Method
The optical density of the cultures was recorded at 600nm, and each absorbance spectrum was normalized to OD600 by division by this value. Background subtraction of spectrophotometer data was performed in Origin 6.1 Software. A ten-point baseline was created by a "positive peak" algorithm then modified to approximate the scattering curve that falls as the inverse fourth power of wavelength.
Results
02 puc pro characterization graph.png

puc promoter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 26
    Illegal XhoI site found at 185
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 574
    Illegal BsaI site found at 583


Design Notes

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Source

PCR applified from plasmid pRKCBC3 provided by Dr. Neil Hunter.

References