Difference between revisions of "Part:BBa K4440000"

 
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VP2C-Tyr1 fusion protein
 
VP2C-Tyr1 fusion protein
  
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===Usage and Biology===
 
===Usage and Biology===
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This part was created for a strategy in which we aimed to concentrate the tyrosinase (Tyr1[BBa_K4440002]) and melanin production into virus-like nanoparticles with the aim to minimize toxicity. For this, the cells expressed viral VP1 protein and the tyrosinase was expressed as a fusion to viral VP2C protein (<b>Figure 1</b>). In this way, VP1 and VP2C multimerize to form a particle, where tyrosinase was localized in the particle.
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[[Image:Figure2 engenering.svg|600px|thumb|center|<b>Figure 1.</b> VP2C-Tyr1, when expressed simultaneously with VP1 protein, is expected to form nanoparticles so that the tyrosinase is directed to the inside of the particles.]]
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In order to produce melanin in nanocompartments this sequence was put under GAL1/10 promoter to produce both parts of the compartment simultaneously.
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We tested the effect of this part in yeast with several experiments: visually the yeast changed color because of a produced pigment (<b>Figure 2</b>), increased cellular fluorescence upon expression of VP2C-Tyr1 was observed (<b>Figure 3</b>).
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[[Image:Vp2c part.svg|600px|thumb|center|<b>Figure 2.</b> Observable color change in VP2C-Tyr1 expressing yeast. ]]
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[[Image:Vp2c flu part.png|600px|thumb|center<b>Figure 3.</b> Microscopy images demonstrating an increase in cell fluorescence upon VP2C-Tyr1 expression, indicative of the presence of some pigment.]]
  
 
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Revision as of 03:00, 12 October 2022


VP2C-Tyr1

VP2C-Tyr1 fusion protein

Usage and Biology

This part was created for a strategy in which we aimed to concentrate the tyrosinase (Tyr1[BBa_K4440002]) and melanin production into virus-like nanoparticles with the aim to minimize toxicity. For this, the cells expressed viral VP1 protein and the tyrosinase was expressed as a fusion to viral VP2C protein (Figure 1). In this way, VP1 and VP2C multimerize to form a particle, where tyrosinase was localized in the particle.

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Figure 1. VP2C-Tyr1, when expressed simultaneously with VP1 protein, is expected to form nanoparticles so that the tyrosinase is directed to the inside of the particles.

In order to produce melanin in nanocompartments this sequence was put under GAL1/10 promoter to produce both parts of the compartment simultaneously.

We tested the effect of this part in yeast with several experiments: visually the yeast changed color because of a produced pigment (Figure 2), increased cellular fluorescence upon expression of VP2C-Tyr1 was observed (Figure 3).

Error creating thumbnail: File missing
Figure 2. Observable color change in VP2C-Tyr1 expressing yeast.
centerFigure 3. Microscopy images demonstrating an increase in cell fluorescence upon VP2C-Tyr1 expression, indicative of the presence of some pigment.

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]