Coding

Part:BBa_K1323009

Designed by: Julia Manalil   Group: iGEM14_Waterloo   (2014-10-04)

DsRed (RFP) Coding Sequence

This is the coding sequence of DsRed_T3 (DNT), a highly fluorescent mutant version of a red fluorescent protein originally found in Discosoma (corals) (Dunn et. al., 2006). It was codon optimized for Staphylococcus aureus by Bose et. al. (2013) and we had this part synthesized by Bio Basic Canada Inc.


(A)DsRED.png (B)DsRED!.tif

DsRed fluorescence was analyzed in both E. coli and S. epidermidis. The DsRed cassette (BBa_K1323015) was incorporated into a pUC57 plasmid containing an erythromycin resistance gene (BBa_K1323011) and Staphylococcal oriV (BBa_K1323018). This plasmid was transformed into the strains using heat shock for E. coli and electroporation for S. epidermidis. Pellets from 50 ml overnight cultures were collected into 1.5 ml eppendorf tubes. To the naked eye (A) E. coli has a pink color (left), whereas in S. epidermidis (middle), a slight pink colour could only be noticed if compared to S. epidermidis without the construct (right). When seen under a RFP filter (B) a high degree of red fluorescence was observed in E. coli (left) and a lower degree of red fluorescence was seen in S. epidermidis (middle). The S. epidermidis negative control (right) showed no observable fluorescence.

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]


References

Bose, J. L., Fey, P. D., and Bayles, K. W. (2013). Genetic Tools to Enhance the Study of Gene Function and Regulation in Staphylococcus aureus. Applied and Environmental Microbiology. 79 (7): 2218-2224.

Dunn, A. K., Millikan, D. S., Adin, D. M., Bose, J. B., and Stabb, E. V. (2006). New rfp- and pES213-Derived Tools for Analyzing Symbiotic Vibrio fisheri Reveal Patterns of Infection and lux Expression In Situ. Applied and Environmental Microbiology. 72 (1): 802-810.


[edit]
Categories
//cds/reporter/rfp
//function/reporter/fluorescence
Parameters
colorRed