Difference between revisions of "Part:BBa K1875003"
 
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==<big>''' CONTRIBUTION '''</big>==
 
==<big>''' CONTRIBUTION '''</big>==
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''' [http://2017.igem.org/Team:Kyoto  iGEM Kyoto 2017] '''  <big>improved</big> this part by connecting this part with ''' [https://parts.igem.org/Part:BBa_K1875003 BBa_K1875003(EGFP)] ''' and added further functions. In this improvement, we obtained a new part ''' [https://parts.igem.org/Part:BBa_K2403003  BBa_K2403003] '''.
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''' [http://2017.igem.org/Team:Kyoto  iGEM Kyoto 2017] '''  <big>improved</big> this part by connecting it with ''' [https://parts.igem.org/Part:BBa_K530008 BBa_K530008 (TDH3 promoter)] ''', adding further functions. In this improvement, we obtained a new part ''' [https://parts.igem.org/Part:BBa_K2403003  BBa_K2403003] '''.
  
 
===Combination with BBa_K530008 (TDH3 promoter)===
 
===Combination with BBa_K530008 (TDH3 promoter)===
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With this combination, this part ''' [https://parts.igem.org/Part:BBa_K1875003  BBa_K1875003(EGFP)] ''' was used to create the part ''' [https://parts.igem.org/Part:BBa_K2403003  BBa_K2403003] ''' which codes EGFP emitting strong green fluorescence. As a result, it was possible to confirm yeast emitting a strong green fluorescence under the microscope (photo or link). In addition, we confirmed that the pine wood nematode eats <i>S. cerevisiae</i>. Such usage is innovative and has greatly contributed to our project.
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This combination gave this part the function as a reporter in <i>S. cerevisiae</i>. Furthermore, by culturing pine wood nematodes with yeast expressing EGFP, feeding on yeast by pine wood nematodes observed, demonstrating an innovative new use and functional contribution of the original parts.
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==<big>''' Information '''</big>==
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Provided by PuiChing-Macau 2023. This team used this mammalian codon optimized EGFP to make a FRET pair (BBa_K4814006). ''' [https://parts.igem.org/Part:BBa_K4814006] '''
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A research (Yang, T. T., Cheng, L., & Kain, S. R., 1996) combined a mutant of GFP with a significantly larger extinction coefficient for excitation at 488 nm with a re-engineered GFP gene sequence containing codons preferentially found in highly expressed human proteins. This improvement increased the fluorescence intensity and higher expression levels, results in higher sensitivity.  
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Sequence derived from: https://www.snapgene.com/plasmids/fluorescent_protein_genes_and_plasmids/EGFP
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Yang, T. T., Cheng, L., & Kain, S. R. (1996). Optimized codon usage and chromophore mutations provide enhanced sensitivity with the green fluorescent protein. Nucleic acids research, 24(22), 4592–4593. https://doi.org/10.1093/nar/24.22.4592

Latest revision as of 13:39, 28 September 2023


This part contains a GFP reporter

This GFP is used in all of the following plasmids: BBa_K1875013, BBa_K1875014, BBa_K1875015, BBa_K1875016, BBa_K1875017, BBa_K1875018, BBa_K1875019.

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]


CONTRIBUTION

[http://2017.igem.org/Team:Kyoto iGEM Kyoto 2017] improved this part by connecting it with BBa_K530008 (TDH3 promoter) , adding further functions. In this improvement, we obtained a new part BBa_K2403003 .

Combination with BBa_K530008 (TDH3 promoter)

This combination gave this part the function as a reporter in S. cerevisiae. Furthermore, by culturing pine wood nematodes with yeast expressing EGFP, feeding on yeast by pine wood nematodes observed, demonstrating an innovative new use and functional contribution of the original parts.

Information

Provided by PuiChing-Macau 2023. This team used this mammalian codon optimized EGFP to make a FRET pair (BBa_K4814006). [1]

A research (Yang, T. T., Cheng, L., & Kain, S. R., 1996) combined a mutant of GFP with a significantly larger extinction coefficient for excitation at 488 nm with a re-engineered GFP gene sequence containing codons preferentially found in highly expressed human proteins. This improvement increased the fluorescence intensity and higher expression levels, results in higher sensitivity.

Sequence derived from: https://www.snapgene.com/plasmids/fluorescent_protein_genes_and_plasmids/EGFP

Yang, T. T., Cheng, L., & Kain, S. R. (1996). Optimized codon usage and chromophore mutations provide enhanced sensitivity with the green fluorescent protein. Nucleic acids research, 24(22), 4592–4593. https://doi.org/10.1093/nar/24.22.4592