Difference between revisions of "Part:BBa K1462090:Design"
 
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===Design Notes===
 
===Design Notes===
  
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To test the localization function of CoxVI, the C-terminus of CoxVI was fused to GFP. Besides,we use MitoTracker Red®CMXRos to stain the mitochondria and do an overlap with GFP. By observing the coefficient value, we can judge that if our leading peptide works.<br/>We adopted fluorescence test to have a visible look at where the fusion protein is located. The following microscope images were obtained. It was observed that green fluorescence concentrated in mitochondria stained by a specific stain, instead of diffusing in the cytosol.The reult proves that CoxVI can locate GFP into the mitochondrial matrix.
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To verify the localization function of CoxVI, the C-terminus of CoxVI was fused with GFP. We adopted fluorescence test to have a visible look at where the GFP is located. The following microscope images were captured(Figure 1). We can observe that the green fluorescence concentrates in a specific area , instead of diffusing homogeneously in the cytosol.The reult proves that CoxVI can perform its function properly
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[[File:G6GT2.jpg|100px|left|frame|Figue1.GFP is targed to the mitochondria]]
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[[File:G6GT.jpg|200px|left|frame|<b>Figure 1.COXVI function verification</b> GFP fused with COXVI is concentrated in a specific area of the cytoplasm,while GFP without COXVI diffuses in the cytoplasm of the cell. ]]
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===Source===
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COXVI is obtained from genome of saccharomyces cerevisiae, the other parts is from the kit provided by official
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===References===
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Ammy C. Maarse1 et al, Subunit IV of yeast cytochrome c oxidase: cloning and nuicleotide sequencing of the gene and partial amino acid sequencing of the mature protein,The EMBO Journal vol.3 no.12 pp.2831 -2837, 1984
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Latest revision as of 13:27, 17 October 2014

Gal1+cox6+GFP+ADH1


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 150
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1325


Design Notes

To verify the localization function of CoxVI, the C-terminus of CoxVI was fused with GFP. We adopted fluorescence test to have a visible look at where the GFP is located. The following microscope images were captured(Figure 1). We can observe that the green fluorescence concentrates in a specific area , instead of diffusing homogeneously in the cytosol.The reult proves that CoxVI can perform its function properly

Figure 1.COXVI function verification GFP fused with COXVI is concentrated in a specific area of the cytoplasm,while GFP without COXVI diffuses in the cytoplasm of the cell.