Difference between revisions of "Part:BBa K2314831"

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"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.  
 
"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]
 
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(BBa_K) to construct a minimal genetic regulatory element “MINI-GRE“ combination. We also selected a commonly used promoter CYC1 and terminator CYC1, and contrusted four circuits to measure the performance of MINI-GRE combination.  
+
We used Pmini promoter with synthetic terminator Tmini(BBa_K2314608) to construct a minimal genetic regulatory element “MINI-GRE“ combination. We also selected a commonly used promoter CYC1 and terminator CYC1, and contrusted four circuits to measure the performance of MINI-GRE combination.  
  
 
[1]Redden H,Alper HS,The development and characterization of synthetic minimal yeast promoters[J],Nature Communication,2015,6 : 7810
 
[1]Redden H,Alper HS,The development and characterization of synthetic minimal yeast promoters[J],Nature Communication,2015,6 : 7810

Revision as of 15:05, 1 November 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(BBa_K2314608) to construct a minimal genetic regulatory element “MINI-GRE“ combination. We also selected a commonly used promoter CYC1 and terminator CYC1, and contrusted four circuits to measure the performance of MINI-GRE combination.

[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--FLO.jpg

the RT-PCR result of mini system:

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

the structure of pmini:

T--OUC-China--MINIP.jpg


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]