Difference between revisions of "Part:BBa K2442206"
Line 12: Line 12:
  
  
−
[[Image:T-Glasgow-finalmtlrimage.png|450px|thumb|left|'''Figure 1:''' GFP response to the sugar induction of regulatory and or reporter plasmid]]
+
[[Image:T-Glasgow-Variants.png|450px|thumb|left|'''Figure 1:''' GFP response to the sugar induction of regulatory and or reporter plasmid]]
 
   
 
   
  

Revision as of 22:23, 1 November 2017

Pmtle + RBS WT + GFP


This part contains the native pmtlE promoter ligated upstream of GFP (E0040). pmtlE is the intergenic region between mtlR and the mtlE coding region in Pseudomonas Flourescens. GFP is taken from the parts registry E0040. This reporter plasmid was used in conjunction with a regulatory plasmid in order to induce the pmtlE.

Characterization

In order to test the working of our reporter constructs, these were ligated into E.coli, DH5a cells. We studied the indiction of this promoter with mtlR with a number of different sugars, mannitol, sorbitol, sucrose, xylose, ribose, and fructose. The reporter plasmid was tested alone with these sugars as well as being tested alongside the regulatory plasmid.


File:T-Glasgow-Variants.png
Figure 1: GFP response to the sugar induction of regulatory and or reporter plasmid


Our results show a fluoresence response both from the reporter plasmid alone as well as in addition with the regulatory plasmid. This suggests that mtlE within this construct is constitutively active. However, upon addition with the regulatory plasmid, there is almost a two fold increase in GFP response, suggesting the regulatory protein still contributes to the induction of pmtlE.


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 829