Part:BBa_K2442206
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.
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 829
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