Difference between revisions of "Part:BBa K3600000"

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<partinfo>BBa_K3600000 short</partinfo>
 
<partinfo>BBa_K3600000 short</partinfo>
  
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This part contains the promoter of HSP12, a heat-shock protein in Saccharomyces Cerevisiae that is activated by the high-osmolarity glycerol (HOG) pathway.<html><a href="#varela1995"><sup>1</sup></a></html>
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HSP12 promoter part plasmid was made in order to create a part plasmid which contains the HSP12 promoter and is compatible with the Yeast Toolkit.
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To create a promoter part plasmid compatible with the yeast toolkit, at first, we designed a gBlock consisting of the 500bp promoter region, that was retrieved from the yeast genome database, and two linking sequences which were added to the ends of the promoter.These linking sequences contained a BsaI and a BsmBI cutting site each to make Golden Gate assembly possible.Then, the gBlock was inserted into the part plasmid entry vector (pYTK001) through a BsmBI assembly.
  
 
===Usage and Biology===
 
===Usage and Biology===
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The HSP12 gene encodes for a heat-shock protein in Saccharomyces Cerevisiae that is activated by the high-osmolarity glycerol (HOG) pathway.<html><a href="#varela1995"><sup>1</sup></a></html>
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* activators: MSN2/4
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The HSP12 promoter is regulated by the transcription factor MSN2/4.
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*HSP12 gene encodes Saccharomyces Cerevisiae's small heat shock proteins.<html><a href="#varela1995"><sup>1</sup></a></html>
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Using the yeast toolkit for modular assembly, <html><a href="#Lee2015"><sup>2</sup></a></html> the HSP12 promoter plasmid was used as a part of type II.
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*The promoter HSP12 has been chosen in order to express fluorescent protein [ref] when the S.cervisiae is exposed to a stress.  
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*The yeast toolkit for modular assembly <html><a href="#Lee2015"><sup>3</sup></a></html> was used to assemble transcriptional units. This promoter was used as a part of type II.
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*The induction of gene transcription is mediated by stress response elements STRE.<html><a href="#Martínez-Pastor1996"><sup>2</sup></a></html>
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Latest revision as of 18:24, 27 October 2020


HSP12 promoter part plasmid

HSP12 promoter part plasmid was made in order to create a part plasmid which contains the HSP12 promoter and is compatible with the Yeast Toolkit. To create a promoter part plasmid compatible with the yeast toolkit, at first, we designed a gBlock consisting of the 500bp promoter region, that was retrieved from the yeast genome database, and two linking sequences which were added to the ends of the promoter.These linking sequences contained a BsaI and a BsmBI cutting site each to make Golden Gate assembly possible.Then, the gBlock was inserted into the part plasmid entry vector (pYTK001) through a BsmBI assembly.

Usage and Biology

The HSP12 gene encodes for a heat-shock protein in Saccharomyces Cerevisiae that is activated by the high-osmolarity glycerol (HOG) pathway.1 The HSP12 promoter is regulated by the transcription factor MSN2/4. Using the yeast toolkit for modular assembly, 2 the HSP12 promoter plasmid was used as a part of type II.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 308
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 326
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 308
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 308
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 57
    Illegal BsaI.rc site found at 578


Measurements

References

[1] Varela, J. C., Praekelt, U. M., Meacock, P. A., Planta, R. J. & Mager, W. H. The Saccharomyces cerevisiae HSP12 gene is activated by the high-osmolarity glycerol pathway and negatively regulated by protein kinase A. Molecular and Cellular Biology 15, 6232–6245 (1995).

[2] Lee, M. E., DeLoache, W. C., Cervantes, B. & Dueber, J. E. A Highly Characterized Yeast Toolkit for Modular, Multipart Assembly. ACS Synth. Biol. 4, 975–986 (2015).

[3] Martínez-Pastor, M. T. et al. The Saccharomyces cerevisiae zinc finger proteins Msn2p and Msn4p are required for transcriptional induction through the stress response element (STRE). The EMBO Journal 15, 2227–2235 (1996).