Not Released
Experience: Works
Not Used
Get This Part
Reporter

Part:BBa_K1323015

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

DsRed Expression Cassette under a Constitutive Promoter

This part is intended for use in Staphylococcal organisms. It is a composite part containing the DsRed coding sequence BBa_K1323009 under the sarA constitutive promoter BBa_K1323021 and translational initiation region BBa_K1323016.


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

DsRed fluorescence was analyzed in both E. coli and S. epidermidis. The DsRed cassette 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]


[edit]
Categories
Parameters
None