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

(→‎Applications of BBa_K1055000)
(→‎Applications of BBa_K1055000)
Line 7: Line 7:
 
''Characterization mKate''
 
''Characterization mKate''
  
−
[[Image:mKate_bac.png|400px|thumb|right|Figure 3. '''Peller of ''E.coli'' Bl21(DE3) cells with expressed mKate. Left side: without UV radiation, right side: with UV radiation ''']]
+
[[Image:mKate_bac.png|400px|thumb|right|Figure 1. '''Peller of ''E.coli'' Bl21(DE3) cells with expressed mKate. Left side: without UV radiation, right side: with UV radiation ''']]
  
  
−
mKate is a red fluorescent protein that we want to use as a acceptor for FRET (Figure 1). According to [http://www.evrogen.com/products/basicFPs.shtml Evrogen] mKate has an excitation maximum at 588 nm and an emission maximum at 633 nm. Figure 2 shows that the excitation maximum is at 579 nm and the emission maximum is 611 nm. The emission maximum is big enough to get stimulated by [https://parts.igem.org/wiki/index.php?title=Part:BBa_K1055001 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.  
+
mKate is a red fluorescent protein that we want to use as a acceptor for FRET (Figure 3). According to [http://www.evrogen.com/products/basicFPs.shtml Evrogen] mKate has an excitation maximum at 588 nm and an emission maximum at 633 nm. Figure 2 shows that the excitation maximum is at 579 nm and the emission maximum is 611 nm. The emission maximum is big enough to get stimulated by [https://parts.igem.org/wiki/index.php?title=Part:BBa_K1055001 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.  
  
  
Line 18: Line 18:
  
  
−
[[Image:MKate-LSSmOrange_FRET_Pair.png|400px|left|thumb|Figure 1. '''Overlay of excitation spectrum (dashed line) and emission spectrum (solid line) of mKate and LSSmOrange ''']]
+
[[Image:MKate-LSSmOrange_FRET_Pair.png|400px|left|thumb|Figure 3. '''Overlay of excitation spectrum (dashed line) and emission spectrum (solid line) of mKate and LSSmOrange ''']]
  
  
Line 56: Line 56:
 
As shown in Figure 4 the expression of mKate is not toxic towards the bacterial host ''E.coli'' Bl21(DE3).
 
As shown in Figure 4 the expression of mKate is not toxic towards the bacterial host ''E.coli'' Bl21(DE3).
  
−
[[Image:Grow_log.png|400px|thumb|Figure 4. '''Semi-logarithmic growth curves of BL21DE3 WT, BL21DE3 mKate and BL21DE3 LSSmOrange. The ln (OD600 values) are plotted against the time [min].''']]
+
[[Image:Grow_log.png|400px|thumb|center|Figure 4. '''Semi-logarithmic growth curves of BL21DE3 WT, BL21DE3 mKate and BL21DE3 LSSmOrange. The ln (OD600 values) are plotted against the time [min].''']]
  
 
===User Reviews===
 
===User Reviews===

Revision as of 22:25, 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_K1055000

Characterization mKate

Figure 1. Peller of E.coli Bl21(DE3) cells with expressed mKate. Left side: without UV radiation, right side: with UV radiation


mKate is a red fluorescent protein that we want to use as a acceptor for FRET (Figure 3). According to [http://www.evrogen.com/products/basicFPs.shtml Evrogen] mKate has an excitation maximum at 588 nm and an emission maximum at 633 nm. Figure 2 shows that the excitation maximum is at 579 nm and the emission maximum is 611 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 2. Excitation spectrum (dashed line) and emission spectrum (solid line) of mKate with marked maximums



Figure 3. Overlay of excitation spectrum (dashed line) and emission spectrum (solid line) of mKate and LSSmOrange


















As shown in Figure 4 the expression of mKate is not toxic towards the bacterial host E.coli Bl21(DE3).

Figure 4. Semi-logarithmic growth curves of BL21DE3 WT, BL21DE3 mKate and BL21DE3 LSSmOrange. The ln (OD600 values) are plotted against the time [min].

User Reviews

UNIQ5d9254a18307a9b6-partinfo-00000000-QINU UNIQ5d9254a18307a9b6-partinfo-00000001-QINU