Difference between revisions of "Part:BBa K1055001:Experience"

(→‎Applications of BBa_K1055001)
(→‎Applications of BBa_K1055001)
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[[Image:mKate-LSSmOrange_FRET_Pair.png|400px|thumb|Figure 1. '''Overlay of exitation spectrum (dashed line) and emission (solid line) of mKate and LSSmOrange''']]
 
  
  
  
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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 [https://parts.igem.org/Part:BBa_K1055000 mKate], the other fluorescent protein (Fig. 1). 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.
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[[Image:LSSmOrange_bac.png|400px|right|thumb|Figure 1. '''Pellet of ''E. coli'' BL21 (DE3) cells with expressed LSSmOrange. Left side without UV radiation, right side with UV radiation''']]
  
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[[Image:LSSmOrange_Diagramm.png|400px|thumb|Figure 2. '''Excitation spectrum (dashed line) and emission spectrum (solid line) of LSSmOrange with marked maximums''']]
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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 [https://parts.igem.org/Part:BBa_K1055000 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.
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[[Image:LSSmOrange_Diagramm.png|center|400px|thumb|Figure 2. '''Excitation spectrum (dashed line) and emission spectrum (solid line) of LSSmOrange with marked maximums''']]
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[[Image:mKate-LSSmOrange_FRET_Pair.png|center|400px|thumb|Figure 3. '''Overlay of exitation spectrum (dashed line) and emission (solid line) of mKate and LSSmOrange''']]
  
  
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As shown in Fig. 4, the expression of LSSmOrange is not toxic towards the bacterial host, ''E. coli'' BL21(DE3).  
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As shown in Fig. 1 and 4, 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:LSSmOrange_bac.png|400px|thumb|Figure 3. '''Cells grown in GFP-medium containing the arabinose promoter regulating GFP expression.''' Induced at time=0h.]]
 
  
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[[Image:Grow_log.png|400px|thumb|Figure 4. '''Cells grown in GFP-medium containing the arabinose promoter regulating GFP expression.''' Induced at time=0h.]]
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[[Image:Grow_log.png|center|400px|thumb|Figure 4. '''Semi log scaled graph of ''E. coli'' BL21(DE3) growth rate expressing either LSSmOrange, mKate or being the wildtype.''']]
  
 
===User Reviews===
 
===User Reviews===

Revision as of 22:29, 4 October 2013

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K1055001

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




As shown in Fig. 1 and 4, 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.

User Reviews

UNIQ8f6b805af5dc7dc1-partinfo-00000000-QINU UNIQ8f6b805af5dc7dc1-partinfo-00000001-QINU