Difference between revisions of "Part:BBa K864680"
 
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Nucleotide sequence of the tetM tetracycline resistance determinant of the streptococcal conjugative shuttle transposon Tn1545.
 
Nucleotide sequence of the tetM tetracycline resistance determinant of the streptococcal conjugative shuttle transposon Tn1545.
 
P Martin, P Trieu-Cuot, and P Courvalin
 
P Martin, P Trieu-Cuot, and P Courvalin
 +
 +
==<strong>OUC-China 2021’s characterization</strong>==
 +
===Design===
 +
We used sfGFP as our reporter gene. When CTE was absent, TetR would bind to the inducible promoter（PI）and prevent RNA polymerase from initiating transcription, thus repressing the expression of reporter gene. If tet was present, TetR would no longer able to bind to the promoter, resulting in the expression of reporter gene.
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===Result===
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https://static.igem.org/mediawiki/parts/thumb/0/01/Ctcs-tetm-sfgfp.PNG/800px-Ctcs-tetm-sfgfp.PNG
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 +
we tested the fluorescence amount produced at different concentrations of tetracycline, and found that it increased the test upper limit compared to the circuit without tetM.
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 19:04, 21 October 2021

tetM

TetM is a tetracycline resistance gene, encoding a ribosomal protection protein. (Martin. et al. 1986)

This gene confers tetracycline resistance (at least 12 µg/ml to E coli Top10 when assembled as BBa_J23110-BBa_B0034-tetM in a middle copy backbone.

Reference

Nucleic Acids Res. 1986 September 11; 14(17): 7047–7058. PMCID: PMC311716 Nucleotide sequence of the tetM tetracycline resistance determinant of the streptococcal conjugative shuttle transposon Tn1545. P Martin, P Trieu-Cuot, and P Courvalin

OUC-China 2021’s characterization

Design

We used sfGFP as our reporter gene. When CTE was absent, TetR would bind to the inducible promoter（PI）and prevent RNA polymerase from initiating transcription, thus repressing the expression of reporter gene. If tet was present, TetR would no longer able to bind to the promoter, resulting in the expression of reporter gene.

Result

800px-Ctcs-tetm-sfgfp.PNG

we tested the fluorescence amount produced at different concentrations of tetracycline, and found that it increased the test upper limit compared to the circuit without tetM.


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 1266