Difference between revisions of "Part:BBa K3257119"

 
 
(One intermediate revision by the same user not shown)
Line 1: Line 1:
−
 
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K3257119 short</partinfo>
 
<partinfo>BBa_K3257119 short</partinfo>
  
 
This part is a truncated mCherry fused two lox sites on both sides. It is used for the verification of the homologous recombination conducted by Cre. We purposely have not added a promoter or terminator in this composite part because, after homologous recombination, a wild-type mCherry will be recombined into this locus and we will be unable to detect red fluorescence since wild-type mCherry cannot be expressed at all.  
 
This part is a truncated mCherry fused two lox sites on both sides. It is used for the verification of the homologous recombination conducted by Cre. We purposely have not added a promoter or terminator in this composite part because, after homologous recombination, a wild-type mCherry will be recombined into this locus and we will be unable to detect red fluorescence since wild-type mCherry cannot be expressed at all.  
 +
 +
'''Prove of well-functional mCherry'''
 +
 +
[[File:MCherry and its mutants.png|center|400px|thumb|'''Figure 1. The relative fluorescence intensity of mCherry and its mutants''' mCherry represents the relative fluorescence intensity of BL21(DE3) transformed with plasmid encoding red fluorescence protein mCherry. The mCherry_Q158X and the mCherry_D159X  represents the relative fluorescence intensity of BL21(DE3) transformed with plasmid encoding nonsense mutated red fluorescence protein mCherry. Significant difference between them shows that mCherry can be expressed and function well inside ''E.coli'' cells.]]
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
Line 9: Line 12:
  
 
<!-- -->
 
<!-- -->
−
<span class='h3bb'>Sequence and Features</span>
+
<span class='h3bb'>'''Sequence and Features'''</span>
 
<partinfo>BBa_K3257119 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3257119 SequenceAndFeatures</partinfo>
  

Latest revision as of 21:25, 21 October 2019

lox5171-truncated mCherry-loxP

This part is a truncated mCherry fused two lox sites on both sides. It is used for the verification of the homologous recombination conducted by Cre. We purposely have not added a promoter or terminator in this composite part because, after homologous recombination, a wild-type mCherry will be recombined into this locus and we will be unable to detect red fluorescence since wild-type mCherry cannot be expressed at all.

Prove of well-functional mCherry

Figure 1. The relative fluorescence intensity of mCherry and its mutants mCherry represents the relative fluorescence intensity of BL21(DE3) transformed with plasmid encoding red fluorescence protein mCherry. The mCherry_Q158X and the mCherry_D159X represents the relative fluorescence intensity of BL21(DE3) transformed with plasmid encoding nonsense mutated red fluorescence protein mCherry. Significant difference between them shows that mCherry can be expressed and function well inside E.coli cells.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 392
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 392
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 392
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 392
  • 1000
    COMPATIBLE WITH RFC[1000]