Difference between revisions of "Part:BBa K2314831"

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the expression level of mini system:
 
the expression level of mini system:
<img src="https://static.igem.org/mediawiki/parts/3/35/T--OUC-China--minifluorescent.png
 
  
"/>
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https://static.igem.org/mediawiki/parts/3/35/T--OUC-China--minifluorescent.png
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the RT-PCR result of mini system:
 
the RT-PCR result of mini system:
<img src="https://static.igem.org/mediawiki/parts/3/35/T--OUC-China--miniRT-PCR.png
 
  
"/>
+
https://static.igem.org/mediawiki/parts/3/35/T--OUC-China--miniRT-PCR.png
 +
 
 
the structure of pmini:
 
the structure of pmini:
<img src="https://static.igem.org/mediawiki/parts/a/a9/T--OUC-China--Pministructure.png
 
  
"/>
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https://static.igem.org/mediawiki/parts/a/a9/T--OUC-China--Pministructure.png
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 03:46, 23 October 2017

Pmini is a very short constitutive promoter in yeast, which is only 116bp, but with a good strength.

"Most of native yeast promoters can stretch hundreds of base pairs. Specifically, a single-gene circuit carrying a 1.5kb gene requires an additional 1kb of regulatory DNA (between the promoter and terminator) for appropriate expression, thus increasing the DNA cargo load by over 60%. However, this problem hasn’t been focused widely. With the development of synthetic biology, creating short but strong promoter is more important. Pmini is a short but strong constitutive promoter in yeast. It consists three main elements. Core element determines the shortest length required for transcription and serves as a platform for hybrid promoter technology. This core element scaffold was built on distinct, essential sequences for promoter function—a TATA box with consensus sequence of TATAWAWR24 followed by a transcription start site (TSS) with consensus sequence of A(Arich)5NYAWNN(Arich)6. UAS element contains transcription factor-binding sites (TFBS) and can aid in RNAP stabilization to enhanced transcription rates. The AT-rich spacer containing 30 nucleotides has a better performance.[1] We used Pmini promoter with synthetic terminator Tmini to construct a “Mini system“ by constructing four circuits. We wanted to compare the expression of “Mini system” with that of the common “CYC1 system” which contains Pcyc1 and Tcyc1.

[1]Redden H,Alper HS,The development and characterization of synthetic minimal yeast promoters[J],Nature Communication,2015,6 : 7810 "


In short,Pmini is a very short constitutive promoter in yeast, which is only 116bp in length, but with strong expression.

the expression level of mini system:

T--OUC-China--minifluorescent.png

the RT-PCR result of mini system:

T--OUC-China--miniRT-PCR.png

the structure of pmini:

T--OUC-China--Pministructure.png


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
    COMPATIBLE WITH RFC[1000]