Difference between revisions of "Part:BBa K227007:Design"
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| − | + | The puc promoter 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 allows 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. | More absorption of light at the LH2 spectra peaks normalized to culture OD corresponds with more transcription and vis versa. | ||
Revision as of 22:30, 18 October 2009
The puc promoter 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 allows 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
puc promoter
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 26
Illegal XhoI site found at 185 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE 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.