Difference between revisions of "Part:BBa K2200007"

 
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GAL4 is a transcription factor of galactose metabolism related genes, which recognizes and binds the promoter UAS through its N-terminal DNA binding domain, and assembles and transcribes the RNA polymerase II complex through its C-terminal activation domain.According to previous studies, it's activity can be up-regulated by adding the P65 sequence.
 
GAL4 is a transcription factor of galactose metabolism related genes, which recognizes and binds the promoter UAS through its N-terminal DNA binding domain, and assembles and transcribes the RNA polymerase II complex through its C-terminal activation domain.According to previous studies, it's activity can be up-regulated by adding the P65 sequence.
  
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Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===
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iGEM_SFLS team improved the https://parts.igem.org/Part:BBa_K1470002 to create a more effecient TAS(transcriptional amplification strategy system)by fusing the NF-κB p65 proteinto the DNA-binding domain of GAL4 protein.The TAS was designed aim to improve the function of Cell-type-specific promoters like tumor-specific promotors.They are able to restrict gene expression to particular cell,but they always weak promoters which can not sufficiently initiate the transcription of downstream genes.As a result,the TAS system is used to enhance the activity of such promoters without loss of cell type specificity.
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K2200007 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2200007 SequenceAndFeatures</partinfo>
  
  
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Uncomment this to enable Functional Parameter display  
 
===Functional Parameters===
 
===Functional Parameters===
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The function of this part was demonstrated by an experiment in a previous paper. CMV promotor was selectively inserted in GAL4-P65,GAL4 and GAL4-VP64 gene.(VP64 was a transcription factor) And GAL4-P65 was also inserted downsream the tumor-specific promotor hTERT. The function of these promotors and transcription factors were measured by the expression of  luciferase in bladder cancer cell lines T24 , 5637, and human human foreskin fibroblasts. The results showed that after adding the sequence of P65, the luciferase activities were obviously up-regulated by the CMV-GAL4-P65 vector group, which means that the GAL4P65 system can significently enhance the expression of downstream gene comparing with the GAL4.
 
<partinfo>BBa_K2200007 parameters</partinfo>
 
<partinfo>BBa_K2200007 parameters</partinfo>
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Revision as of 13:15, 29 October 2017


Gal4 transcription factor is a positive regulator.

GAL4 is a transcription factor of galactose metabolism related genes, which recognizes and binds the promoter UAS through its N-terminal DNA binding domain, and assembles and transcribes the RNA polymerase II complex through its C-terminal activation domain.According to previous studies, it's activity can be up-regulated by adding the P65 sequence. 

Add more about the biology of this part here

Usage and Biology

iGEM_SFLS team improved the https://parts.igem.org/Part:BBa_K1470002 to create a more effecient TAS(transcriptional amplification strategy system)by fusing the NF-κB p65 proteinto the DNA-binding domain of GAL4 protein.The TAS was designed aim to improve the function of Cell-type-specific promoters like tumor-specific promotors.They are able to restrict gene expression to particular cell,but they always weak promoters which can not sufficiently initiate the transcription of downstream genes.As a result,the TAS system is used to enhance the activity of such promoters without loss of cell type specificity.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 710
    Illegal XhoI site found at 218
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 886
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 137


Uncomment this to enable Functional Parameter display

Functional Parameters

The function of this part was demonstrated by an experiment in a previous paper. CMV promotor was selectively inserted in GAL4-P65,GAL4 and GAL4-VP64 gene.(VP64 was a transcription factor) And GAL4-P65 was also inserted downsream the tumor-specific promotor hTERT. The function of these promotors and transcription factors were measured by the expression of luciferase in bladder cancer cell lines T24 , 5637, and human human foreskin fibroblasts. The results showed that after adding the sequence of P65, the luciferase activities were obviously up-regulated by the CMV-GAL4-P65 vector group, which means that the GAL4P65 system can significently enhance the expression of downstream gene comparing with the GAL4.