Difference between revisions of "Part:BBa K1055001"

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Furthermore we did homology modeling with this part, please refer to our [[http://2013.igem.org/Team:TU_Darmstadt/modelling/Structure wiki page]], whose best model results are illustrated below:
 
Furthermore we did homology modeling with this part, please refer to our [[http://2013.igem.org/Team:TU_Darmstadt/modelling/Structure wiki page]], whose best model results are illustrated below:
  
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[[Image:LssmOrange_first_activeHelix.png|center|400px|thumb|Figure 3. '''Overlay of exitation spectrum (dashed line) and emission (solid line) of mKate and LSSmOrange''']]
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[[Image:LssmOrange_first_activeHelix.png|center|400px|thumb|]]
  
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The expression of LSSmOrange is not toxic towards the bacterial host, ''E. coli'' BL21(DE3). The growth rate was in a usual time scale of approximately 30 minutes.
  
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[[Image:Grow_log.png|center|400px|thumb|center|Figure 4. '''Semi log scaled graph of ''E. coli'' BL21(DE3) growth rate expressing either LSSmOrange, mKate or being the wildtype.''']]
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
  

Revision as of 18:08, 12 October 2013

Status: 500 Content-type: text/html

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LssmOrange is a fluorophor derived from dsRed and is a FRET partner of mKate [BBa_K1055000]


Usage and Biology

Figure 1. Pellet of E. coli BL21 (DE3) cells with expressed LSSmOrange. Left side without UV radiation, right side with UV radiation

LSSmOrange is an orange fluorescent protein that we want to use for FRET, we us it as a donor. LSSmOrange has an excitation maximum by 437 nm and an emission maximum by 572 nm. The excitation maximum is at 415 nm, another at 453 nm and our measured emission maximum is at 564 nm (Fig. 2). This aberration changes nothing on the FRET system. The excitation maximum is big enough to stimulate mKate, the other fluorescent protein (Fig. 3). Of course, we also checked the emission and excitation of our E. coli BL21(DE3) cells (data not shown) and we can eliminate the theories that these cells disturb this florescence measurement.

Figure 2. Excitation spectrum (dashed line) and emission spectrum (solid line) of LSSmOrange with marked maximums
Figure 3. Overlay of exitation spectrum (dashed line) and emission (solid line) of mKate and LSSmOrange

Furthermore we did homology modeling with this part, please refer to our http://2013.igem.org/Team:TU_Darmstadt/modelling/Structure wiki page, whose best model results are illustrated below:

LssmOrange first activeHelix.png

The expression of LSSmOrange is not toxic towards the bacterial host, E. coli BL21(DE3). The growth rate was in a usual time scale of approximately 30 minutes.


Figure 4. Semi log scaled graph of E. coli BL21(DE3) growth rate expressing either LSSmOrange, mKate or being the wildtype.

Sequence and Features Status: 500 Content-type: text/html

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