Project

Part:BBa_K404316

Designed by: Freiburg Bioware 2010   Group: iGEM10_Freiburg_Bioware   (2010-10-21)

EGFP

eGPP
BioBrick Nr. BBa_K404316
RFC standard RFC 25
Requirement pSB1C3
Submitted by [http://2010.igem.org/Team:Freiburg_Bioware FreiGEM 2010]









EGFP was improved by the iGEM team Freiburg_Bioware 2010 by converting it into RFC 25 standard.
GFP is a fluorescent protein, consisting of 238 aa (=26,9 kDa), which emits green light after being exposed to blue light. Osamu Shimomura described the first GFP protein derived from Aequorea Victoria in 1962. Its excitation spectrum has two peaks at 396 and 475 nm [Heim et al., 1995]. Due to the second excitation peak at 475 nm, GFP protein is very photo stable [Chalfie et al., 1994]. 
eGFP (=enhanced GFP) is a potential derivate of GFP, which finds broad usage in mammalian cells. Compared to the wild-type protein, eGFP is a codon optimized GFP and is achieves up to 10fold higher expression rate and up to 30 fold higher bioluminescence

 



 


Usage and Biology

eGFP is used as a marker gene for detection of transfected cells, e.g. tumor cells.


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 641

References

Cormack, B.P., Valdivia, R.H. & Falkow, S., 1996. FACS-optimized mutants of the green fluorescent protein (GFP). Gene, 173(1 Spec No), pp.33-38. Available at: http://www.ncbi.nlm.nih.gov/pubmed/8707053.
Heim, R., Cubitt, A.B. & Tsien, R.Y., 1995. Improved green fluorescence. Nature, 373(6516), pp.663-664. Available at: http://www.nature.com/nature/journal/v373/n6516/abs/373663b0.html.
Chalfie M, Tu Y, Euskirchen G, Ward W, Prasher D 1994. "Green fluorescent protein as a marker for gene expression". Science 263 (5148): 802–5

[edit]
Categories
//function/reporter
Parameters
None