Reporter

Part:BBa_K1875003

Designed by: Jeffrey Marano   Group: iGEM16_BostonU   (2016-10-11)


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

[edit]
Categories
Parameters
None