Difference between revisions of "Part:BBa K2611010"
 
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We added a spacer sequence and a NGG site closely before the promoter([https://parts.igem.org/Part:BBa_J23101 BBa_J23101]
 
We added a spacer sequence and a NGG site closely before the promoter([https://parts.igem.org/Part:BBa_J23101 BBa_J23101]
−
). This spacer sequence can be recognized by the sgRNA we designed to direct dCas9. Once dCas9 binds with the spacer sequence, the expression of GFP will be repressed.We have submitted the sgRNA part (BBa_K2611000) and spacer-GFP part(BBa_K2611001). In this composite part, we linked them together.We observed positive colonies transformed with sgRNA(spacer J23101-GFP)-spacer J23101-GFP(part BBa_K2611010) under a stereo fluorescence microscope.Green fluorescence was observed.(Fig.1)
+
). This spacer sequence can be recognized by the sgRNA we designed to direct dCas9. Once dCas9 binds with the spacer sequence, the expression of GFP will be repressed.We have submitted the sgRNA part ([https://parts.igem.org/Part:BBa_K2611000 BBa_K2611000]) and spacer-GFP part([https://parts.igem.org/Part:BBa_K2611001 BBa_K2611001]). In this composite part, we linked them together.We observed positive colonies transformed with this part under a stereo fluorescence microscope.Green fluorescence was observed.(Fig.1) Besides,we also mearsured fluorescent intensity of bacteria (transformed with BBa_K2611010) solutions. (Fig.2)Compared with the NC, transformated bacteria solutions' fluorescence intensity/OD are much higher, which indicates GFP was expressed successfully in our transformated bacteria.
  
 
[[File:Long description.png]]
 
[[File:Long description.png]]
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 +
[[File:BBa_K2611010 Fluorescence.png]]
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 02:18, 18 October 2018


sgRNA(spacer J23101-GFP)-spacer J23101-GFP

We added a spacer sequence and a NGG site closely before the promoter(BBa_J23101 ). This spacer sequence can be recognized by the sgRNA we designed to direct dCas9. Once dCas9 binds with the spacer sequence, the expression of GFP will be repressed.We have submitted the sgRNA part (BBa_K2611000) and spacer-GFP part(BBa_K2611001). In this composite part, we linked them together.We observed positive colonies transformed with this part under a stereo fluorescence microscope.Green fluorescence was observed.(Fig.1) Besides,we also mearsured fluorescent intensity of bacteria (transformed with BBa_K2611010) solutions. (Fig.2)Compared with the NC, transformated bacteria solutions' fluorescence intensity/OD are much higher, which indicates GFP was expressed successfully in our transformated bacteria.

Long description.png

BBa K2611010 Fluorescence.png

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 226
    Illegal NheI site found at 249
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 924