Difference between revisions of "Part:BBa K1493200"

(→‎Ferric regulator Pfri)
(→‎Characterization)
Line 14: Line 14:
  
 
=Characterization=
 
=Characterization=
−
<p>The biobrick was tested in a shuttle plasmid (SEVA) and was coupled behind an IPTG inducible promoter. Plasmid was transformed in <i>P.putida</i> KT2440. Cultures were inoculated in 10ml M9 minimal medium containing iron and were incubated in 30°C while shaken overnight. The next morning it was noted that transformants were greener than other cultures, those being Wildtype KT2400 and <i>P.putida</i> containing an empty plasmid. Pyoverdine was measured using a spectrometer with wavelengths ranging from 350-460nm. A higher peak can be seen for transformants containing an extra gene of <i>Pfri</i>
+
 
−
</p>
+
  
  

Revision as of 09:12, 11 October 2014

Ferric regulator Pfri

Pfri was obtained from Pseudomonas putida KT2440, it functions as a transcription regulator that initiates transcription of genes involved in pyoverdine synthesis. Pyoverdine is a flourescent compound that is produced by P.putida that binds to iron (III). This gene was used in order to try to increase pyoverdine production P.putida Read more on our wiki [http://2014.igem.org/Team:Wageningen_UR]


Cloning

Primers used for cloning were:
Fw: 5'-GTTTCTTCGAATTCGCGGCCGCTTCTAGATGGCGGAACAACTATCCACAAGTAAG-3'
Rev: 5-'GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTATCAGGCCTGGCGACTGGC-3'
note: These primers already include the biobrick preffix and suffix


Charecterization

Characterization

Usage and Biology

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]