Difference between revisions of "Part:BBa K1055000:Experience"

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'''Characterization LssmOrange'''
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'''Characterization mKate'''
  
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This biobrick did measure up to our expectations as shown in the following data. We also used this promoter to express our PET cleaving enzyme pNB-Est13, which is anchored C-terminal to EstA.(''E. Coli'' membrane anchor protein). AraC-Pbad shows a  low background activity and a good respose to the induction with Arabinose.  
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mKate2 is a red fluorescent protein that we want to use as a acceptor for FRET. According to Evrogen [2] mKate2 has an excitation maximum at 588 nm and an emission maximum at 633 nm. The emission maximum is big enough to get stimulated by LSSmOrange, the other fluorescent protein. Of course, we checked also the emission and excitation of our BL21(DE3) cells (data not shown) and we can eliminate the theories that these cells disturb this florescence measurement.  
  
  
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[[Image:Spectra_mKate.png|400px|thumb|Figure 1. '''Cells grown in GFP-medium containing the arabinose promoter regulating GFP expression.''' Induced at time=0h.]]
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[[Image:Spectra_mKate.png|400px|thumb|Figure 1. '''Excitation Spectrum (dashed line) and emission spectrum (solid line) of mKate with marked maximums''' Induced at time=0h.]]
  
 
[[Image:MKate-LSSmOrange_FRET_Pair.png|400px|thumb|Figure 1. '''Cells grown in GFP-medium containing the arabinose promoter regulating GFP expression.''' Induced at time=0h.]]
 
[[Image:MKate-LSSmOrange_FRET_Pair.png|400px|thumb|Figure 1. '''Cells grown in GFP-medium containing the arabinose promoter regulating GFP expression.''' Induced at time=0h.]]

Revision as of 21:51, 4 October 2013

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Characterization mKate

mKate2 is a red fluorescent protein that we want to use as a acceptor for FRET. According to Evrogen [2] mKate2 has an excitation maximum at 588 nm and an emission maximum at 633 nm. The emission maximum is big enough to get stimulated by LSSmOrange, the other fluorescent protein. Of course, we checked also the emission and excitation of our BL21(DE3) cells (data not shown) and we can eliminate the theories that these cells disturb this florescence measurement.


Figure 1. Excitation Spectrum (dashed line) and emission spectrum (solid line) of mKate with marked maximums Induced at time=0h.
Figure 1. Cells grown in GFP-medium containing the arabinose promoter regulating GFP expression. Induced at time=0h.
Figure 1. Cells grown in GFP-medium containing the arabinose promoter regulating GFP expression. Induced at time=0h.
Figure 1. Cells grown in GFP-medium containing the arabinose promoter regulating GFP expression. Induced at time=0h.