Difference between revisions of "Part:BBa K1323015"

 
 
(6 intermediate revisions by 2 users not shown)
Line 1: Line 1:
 
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K1323015 short</partinfo>
 
<partinfo>BBa_K1323015 short</partinfo>
  
TBA
+
This part is intended for use in Staphylococcal organisms. It is a composite part containing the DsRed coding sequence [https://parts.igem.org/Part:BBa_K1323009 BBa_K1323009] under the ''sarA'' constitutive promoter [https://parts.igem.org/Part:BBa_K1323021 BBa_K1323021] and  translational initiation region [https://parts.igem.org/Part:BBa_K1323016 BBa_K1323016].
 +
 
 +
 
 +
(A)[[File:DsRED.png]]    (B)[[File: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 ([https://parts.igem.org/Part:BBa_K1323011 BBa_K1323011]) and Staphylococcal oriV ([https://parts.igem.org/Part:BBa_K1323018 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.
 +
 
 +
 
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 15:51, 2 November 2014

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]