Difference between revisions of "Part:BBa K792002"

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'''Contribution Summary:'''
 
'''Contribution Summary:'''
  
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Team NUS_Singapore 2021 uccessfully expressed GFP using this part to quantify the performance. Characterization of part performance was also done using a GFP reporter. Check NUS Singapore 2021 wiki to view more information: https://2021.igem.org/Team:NUS_Singapore/Results  
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Team NUS_Singapore 2021 has successfully expressed GFP using this part to quantify the performance. Characterization of part performance was also done using a GFP reporter. Check NUS Singapore 2021 wiki to view more information: https://2021.igem.org/Team:NUS_Singapore/Results  
  
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[[File:T--NUS_Singapore--results_tab1_2.png|700px|thumb|center|Figure 1: Image of the media supernatant, resuspended cells and blank control media under gel visualizer blue light, media and supernatant are clearly fluorescent compared to the control media.]]
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[[File:T--NUS Singapore--GFP plateimage.png|700px|thumb|center|Figure 1: Image of the media supernatant, resuspended cells and blank control media under gel visualizer blue light, media and supernatant are clearly fluorescent compared to the control media.]]
  
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[[File:T--NUS_Singapore--results_tab1_1.png|800px|thumb|center|Figure 2: mFa-GFP Assembly schematic of the construct used to characterize performance of this secretion tag.]]
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This part was placed at the N-terminus of a GFP reporter gene. The expression of the mFa-GFP fusion protein was controlled using a Gal1 inducible promoter (Figure 2).
  
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[[File:T--NUS_Singapore--results_tab1_3.png|800px|thumb|center|Figure 3: GFP absorbance of the supernatant and cells of both induced and uninduced pGmFaGFP-H (BY4741), after normalization to blank media. Induced cells secreted approximately 42% of the GFP, while uninduced cells produced and secreted significantly less GFP than induced cells.]]
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[[File:T--NUS Singapore--NUS Singapore--pGmGFP-H.png|800px|thumb|center|Figure 2: mFa-GFP Assembly schematic of the construct used to characterize performance of this secretion tag.]]
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GFP absorbance of the supernatant and cells of both induced and uninduced pGmFaGFP-H (BY4741) was recorded (figure 3) after 24 hrs of induction in galactose enriched yeast growth medium. Secretion efficacy of the GFP reporter gene was approximated to about 42%.
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[[File:T--NUS Singapore--pGmGFP-H graph.png|800px|thumb|center|Figure 3: GFP absorbance of the supernatant and cells of both induced and uninduced pGmFaGFP-H (BY4741), after normalization to blank media. Induced cells secreted approximately 42% of the GFP, while uninduced cells produced and secreted significantly less GFP than induced cells.]]
  
 
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Revision as of 20:43, 19 October 2021

Secretion tag from yeast α-factor mating pheromone (MFα1)

This part is the secretion signal peptide from the yeast α-mating factor. This signal peptide directs the secretion of the produced protein, and therefore allows the exportation of it. This peptides are cleaved once the protein is in the lumen of the ER.


To use this tag, attached it directly to your proteins CDS.


See also: A Kozak sequence from α-factor mating pheromone (MFα1) BBa_K792001
This parts was originally built to be used in [http://2012.igem.org/Team:Buenos_Aires iGEM BsAs 2012 project] to implement a cross-feeding regulatory system between two different yeast strains with specific auxotrophy. [http://2012.igem.org/Team:Buenos_Aires Visit our wiki] to read details about the designing process and implementation details of our system. Also feel free to contact us for any question regarding this part. Igem.bsas.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]

Part Contributions

NUS Singapore 2021

Authors:

Tania Santosh Nair, Chew Chin Wei, Linus Tan

Contribution Summary:

Team NUS_Singapore 2021 has successfully expressed GFP using this part to quantify the performance. Characterization of part performance was also done using a GFP reporter. Check NUS Singapore 2021 wiki to view more information: https://2021.igem.org/Team:NUS_Singapore/Results

Figure 1: Image of the media supernatant, resuspended cells and blank control media under gel visualizer blue light, media and supernatant are clearly fluorescent compared to the control media.

This part was placed at the N-terminus of a GFP reporter gene. The expression of the mFa-GFP fusion protein was controlled using a Gal1 inducible promoter (Figure 2).

Figure 2: mFa-GFP Assembly schematic of the construct used to characterize performance of this secretion tag.

GFP absorbance of the supernatant and cells of both induced and uninduced pGmFaGFP-H (BY4741) was recorded (figure 3) after 24 hrs of induction in galactose enriched yeast growth medium. Secretion efficacy of the GFP reporter gene was approximated to about 42%.

Figure 3: GFP absorbance of the supernatant and cells of both induced and uninduced pGmFaGFP-H (BY4741), after normalization to blank media. Induced cells secreted approximately 42% of the GFP, while uninduced cells produced and secreted significantly less GFP than induced cells.


Additional Information

Please visit our results page for more information on the characterization data of this part here: https://2021.igem.org/Team:NUS_Singapore/Results