Difference between revisions of "Part:BBa K592022"

 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K592022 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K592022 SequenceAndFeatures</partinfo>
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===Experimental Validation===
  
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2017 Team NYU Abu Dhabi attempted to express cjBlue in DH5-α using composite part BBa_K592022. Our assembly and transformation [http://2017.igem.org/Team:NYU_Abu_Dhabi/Protocols protocols] were followed in order to obtain the part. All initial, intermediate, and final products obtained throughout the 3A assembly were digested with EcoRI-HF following our digestion [http://2017.igem.org/Team:NYU_Abu_Dhabi/Protocols protocol] and run in a 1.2% agarose gel to confirm that the constructs were of the right size (Figure 1).
  
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"https://static.igem.org/mediawiki/2017/a/a9/Gel_Result_cjBlue_NYUAD2017.png"
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However, no blue colonies were observed after 24 hs of incubation in agar and no blue pellet was obtained after centrifuging an overnight LB broth culture at 7000 rpm for 5 min. The reason for the lack of expression remains elusive.
 
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===Functional Parameters===
 
===Functional Parameters===
 
<partinfo>BBa_K592022 parameters</partinfo>
 
<partinfo>BBa_K592022 parameters</partinfo>
 
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Latest revision as of 22:13, 27 October 2017

cjBlue Green Coding Device

This part is constitutively expressed BBa_K592011, the cjBlue chromoprotein. The color development is slow, it is readily visible to naked eye after 24-48 hours of incubation.

CjBlue plate small.jpg

Escherichia coli constitutively expressing cjBlue (BBa_K592011).


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Experimental Validation

2017 Team NYU Abu Dhabi attempted to express cjBlue in DH5-α using composite part BBa_K592022. Our assembly and transformation [http://2017.igem.org/Team:NYU_Abu_Dhabi/Protocols protocols] were followed in order to obtain the part. All initial, intermediate, and final products obtained throughout the 3A assembly were digested with EcoRI-HF following our digestion [http://2017.igem.org/Team:NYU_Abu_Dhabi/Protocols protocol] and run in a 1.2% agarose gel to confirm that the constructs were of the right size (Figure 1).

"Gel_Result_cjBlue_NYUAD2017.png"

However, no blue colonies were observed after 24 hs of incubation in agar and no blue pellet was obtained after centrifuging an overnight LB broth culture at 7000 rpm for 5 min. The reason for the lack of expression remains elusive.