Difference between revisions of "Part:BBa K4158010"
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We demonstrated that this new part could detect progesterone in the cell-free protein synthesis system.  
 
We demonstrated that this new part could detect progesterone in the cell-free protein synthesis system.  
  
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[[File:Waseda Tokyo result of progesterone sensing by SRTF1.png|500px|thumb|center|Fig. 1. result of progesterone sensing by SRTF1]]
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[[File:Waseda Tokyo gene circuit of progesterone detector.png|500px|thumb|center|Fig. 1. gene circuit of progesterone detector]]
  
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<b>Fig. 1.</b> shows the result of the cell-free protein synthesis reaction. All of the three samples contain the cell-free extracts expressing the transcription factor SRTF1. We added progesterone as 100uM in final concentration. 
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[[File:Waseda Tokyo result of progesterone sensing by SRTF1.png|500px|thumb|center|Fig. 2. result of progesterone sensing by SRTF1]]
  
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We could confirm below from <b>Fig. 1.</b>.  
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<b>Fig. 2.</b> shows the result of the cell-free protein synthesis reaction. All of the three samples contain the cell-free extracts expressing the transcription factor SRTF1. We added progesterone as 100uM in final concentration. 
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We could confirm below from <b>Fig. 2.</b>.  
  
 
<ul>  
 
<ul>  

Revision as of 11:29, 10 October 2022


Psrtf1-GFP

This part contains RBS, GFPuv coding site and a promoter regulated by SRTF1 and Progesterone((1S,3aS,3bS,9aR,9bS,11aS)-1-Acetyl-9a,11a-dimethyl-1,2,3,3a,3b,4,5,8,9,9a,9b,10,11,11a-tetradecahydro-7H-cyclopenta[a]phenanthren-7-one) and works as the reporter plasmid to confirm exist of Progesterone and SRTF1.

RBS+GFPuv sequence is the same as BBa_K4158011.

We designed Psrtf-GFP(BBa_K4158010), and confirmed its activity in vitro. This part is the reporter plasmid used for progesterone detection. It encodes GFP gene in the downstream of the binding site of SRTF1, the transcriptional factor specific to progesterone. 

Promoter and SRTF1 binding site: We cited information about the sequence of SRTF1 dependent promoter from paper[1].

Consequently, the promoter sequence was the same as No part name specified with partinfo tag.) was successfully achieved</b>. we achieved a project based on BioBricks, which is an important standard component in synthetic biology. </li> </ul>


Thus, we succeeded in engineering SRTF1 regulated gfp reporter plasmid(BBa_K4158010). (For more details, go to https://2022.igem.wiki/waseda-tokyo/results#progesterone-detection.)

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 13
    Illegal NheI site found at 36
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 826
    Illegal XhoI site found at 529
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


References

1. Sankar K et al. A progesterone biosensor derived from microbial screening. ACS Sens. 7(4):1132-1137(2022).