Difference between revisions of "Part:BBa K322246:Design"

 
(→‎Design Notes)
 
(One intermediate revision by the same user not shown)
Line 1: Line 1:
−
 
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K322246 short</partinfo>
 
<partinfo>BBa_K322246 short</partinfo>
Line 7: Line 6:
  
 
===Design Notes===
 
===Design Notes===
−
Sequence date: initial introduction of this mutation into K216015 (with PyeaR promoter and RBS) failed (sequence of 123 Aug 2010) and was repeated by Will and Lu. Positive results confirming the mutation were obtained by Lu for 9 of 14 clones on 7 Sep 2010. The mutated coding sequence was then amplified and cloned in pSB1C3. Sequence confirmation 22 Sep 2010; forward sequence is fine, reverse sequence good but missing the starting T of the suffix - goes straight from TAA to ACTAGT rather than TAATACTAGT. Compared to other mistakes in submitted BioBricks this is a pretty minor one, but still annoying.
 
−
 
−
 
−
 
  
 +
This luciferase seems to be temperature sensitive, and growing the host at 30°C rather than 37°C considerably improves brightness.
  
 
===Source===
 
===Source===
Line 18: Line 14:
  
 
===References===
 
===References===
 +
 +
Shapiro, E., Lu, C. & Baneyx, F. (2005). A set of multicolored Photinus pyralis luciferase mutants for in vivo bioluminescence applications. Protein Engineering, Design and Selection 18, 581-587.

Latest revision as of 19:22, 7 October 2010

Red emitting firefly luciferase mutant (S284T)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 808


Design Notes

This luciferase seems to be temperature sensitive, and growing the host at 30°C rather than 37°C considerably improves brightness.

Source

Photinus pyralis

References

Shapiro, E., Lu, C. & Baneyx, F. (2005). A set of multicolored Photinus pyralis luciferase mutants for in vivo bioluminescence applications. Protein Engineering, Design and Selection 18, 581-587.