Difference between revisions of "Part:BBa K2557050"

 
 
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TetR is a repressor that binds to TetO sequences and inhibits the expression of downstream genes. iGEM17_Oxford inserted a TEV cleavage site in TetR (BBa_K2450202), so TetR can be recognized and cleaved by TEV protease, losing binding function and releasing inhibition of downstream genes. In our system, we use this TetR as a negative regulatory protein of the recombinase.
 
TetR is a repressor that binds to TetO sequences and inhibits the expression of downstream genes. iGEM17_Oxford inserted a TEV cleavage site in TetR (BBa_K2450202), so TetR can be recognized and cleaved by TEV protease, losing binding function and releasing inhibition of downstream genes. In our system, we use this TetR as a negative regulatory protein of the recombinase.
  
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===Usage and Biology===
 
===Usage and Biology===
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<html>
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<img src="https://static.igem.org/mediawiki/parts/d/db/T--NAU-CHINA-liuzhang_lvse.jpg"width="600"/>
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</html>
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Fig.1 Inhibition of tetR on promoter with tetO sequence
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(A)Fluorescence microscope observation of HEK 293T only transfect with plasmids containing promoters with tetO sequence.
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(B)Fluorescence microscope observation of HEK 293T transfect with plasmids containing promoters with tetO sequence and tetR.
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Fig. 1 shows that TetR has obvious inhibitory effects on the three promoters downstream of TetO.
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===Improvement===
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This part was modified from BBa_K2450202, which we codon optimized and characterized in HEK 293T.
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===References===
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1. Ramos, J. L. et al. The TetR Family of Transcriptional Repressors The TetR Family of Transcriptional Repressors. Microbiol. Mol. Biol. Rev. 69, 326–356 (2005).
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K2557050 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2557050 SequenceAndFeatures</partinfo>

Latest revision as of 17:26, 17 October 2018


TetR with TEV cleavage site

TetR is a repressor that binds to TetO sequences and inhibits the expression of downstream genes. iGEM17_Oxford inserted a TEV cleavage site in TetR (BBa_K2450202), so TetR can be recognized and cleaved by TEV protease, losing binding function and releasing inhibition of downstream genes. In our system, we use this TetR as a negative regulatory protein of the recombinase.


Usage and Biology

Fig.1 Inhibition of tetR on promoter with tetO sequence (A)Fluorescence microscope observation of HEK 293T only transfect with plasmids containing promoters with tetO sequence. (B)Fluorescence microscope observation of HEK 293T transfect with plasmids containing promoters with tetO sequence and tetR.

Fig. 1 shows that TetR has obvious inhibitory effects on the three promoters downstream of TetO.


Improvement

This part was modified from BBa_K2450202, which we codon optimized and characterized in HEK 293T.


References

1. Ramos, J. L. et al. The TetR Family of Transcriptional Repressors The TetR Family of Transcriptional Repressors. Microbiol. Mol. Biol. Rev. 69, 326–356 (2005).


Sequence and Features


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 BsaI.rc site found at 361