Difference between revisions of "Part:BBa K1031510"
 
Line 3: Line 3:
 
<html>
 
<html>
 
<p>For detailed information concerning HpaR, please visit <a href="http://2013.igem.org/Team:Peking/Project/BioSensors/HcaR#Mileston2">2013 Peking iGEM Biosensor HpaR</a></p>
 
<p>For detailed information concerning HpaR, please visit <a href="http://2013.igem.org/Team:Peking/Project/BioSensors/HcaR#Mileston2">2013 Peking iGEM Biosensor HpaR</a></p>
 +
 +
<img src="https://static.igem.org/mediawiki/igem.org/c/c9/Peking_Logo.jpg" style="width:960px;"/>
 
</html>
 
</html>
  
Line 11: Line 13:
  
 
<html>
 
<html>
−
<img src="https://static.igem.org/mediawiki/igem.org/8/81/Peking2013_HpaRFig4.jpg", width=600px; />
+
<img src="https://static.igem.org/mediawiki/igem.org/8/81/Peking2013_HpaRFig4.jpg" style="width:600px; margin-left:170px"  />
 +
<p style="text-align:center"><b>Fig 1</b>' The hpa cluster map in genome. The arrows inside the squares show the transcriptional direction of genes.Pr, Pg, Px, Pa and Pbc, represent the promoter controlling cognate clusters. Addition of 3-hydroxyphenylacetic acid can derepress the promoters inhibited by HpaR (for Pr and Pg activation) or HpaA (for Pbc activation). Finally the enzymes in these clusters will degrade the 3-HPAA to the intermediates in TCA cycle.
 
</html>
 
</html>
−
 
−
'''Fig 1''' The hpa cluster map in genome. The arrows inside the squares show the transcriptional direction of genes.Pr, Pg, Px, Pa and Pbc, represent the promoter controlling cognate clusters. Addition of 3-hydroxyphenylacetic acid can derepress the promoters inhibited by HpaR (for Pr and Pg activation) or HpaA (for Pbc activation). Finally the enzymes in these clusters will degrade the 3-HPAA to the intermediates in TCA cycle.
 
−
 
  
 
hpa cluster consists of three operons. The regulator gene, hpaR, is transcribed in the divert direction to other genes under Pr promoter. The adjacent promoter, Pg, initiates the transcription of the functional hpaGEDFHI operon. Pr and Pg are both regulated by HpaR and located in the intergenic region between the hpaR and hpaG ('''Fig.1''').  
 
hpa cluster consists of three operons. The regulator gene, hpaR, is transcribed in the divert direction to other genes under Pr promoter. The adjacent promoter, Pg, initiates the transcription of the functional hpaGEDFHI operon. Pr and Pg are both regulated by HpaR and located in the intergenic region between the hpaR and hpaG ('''Fig.1''').  
Line 21: Line 21:
  
 
<html>
 
<html>
−
<img src="https://static.igem.org/mediawiki/igem.org/b/bb/Peking2013_HpaRFig3.jpg", width=600px; />
+
<img src="https://static.igem.org/mediawiki/igem.org/b/bb/Peking2013_HpaRFig3.jpg" style="width:600px; margin-left:170px" />
 +
<p style="text-align:center"><b>Fig 2</b> The degradation pathway of 3HPAA. The letters on the arrows are the names of genes in the clusters. eg. HpaB,C means the step 1 is catalyzed by the product of hpaB and hpaC.
 
</html>
 
</html>
−
 
−
'''Fig 2''' The degradation pathway of 3HPAA. The letters on the arrows are the names of genes in the clusters. eg. HpaB,C means the step 1 is catalyzed by the product of hpaB and hpaC.
 
  
  
Line 42: Line 41:
  
 
<html>
 
<html>
−
<img src="https://static.igem.org/mediawiki/igem.org/thumb/7/7a/Peking2013_part_HpaR-TT.png/800px-Peking2013_part_HpaR-TT.png", width=600px; />
+
<img src="https://static.igem.org/mediawiki/igem.org/thumb/7/7a/Peking2013_part_HpaR-TT.png/800px-Peking2013_part_HpaR-TT.png" style="width:600px; margin-left:170px"  />
 +
<p style="text-align:center"><b>Fig 3</b>' Schematic diagram of HpaR biosensor circuit. Orange arrowheads represent promoters. RBS is shown as green oval and red square stands for B0015 terminator.
 
</html>
 
</html>
−
 
−
'''Fig 3''' Schematic diagram of HpaR biosensor circuit. Orange arrowheads represent promoters. RBS is shown as green oval and red square stands for B0015 terminator.
 
  
 
   
 
   

Latest revision as of 22:09, 27 September 2013

HpaR-Terminator

For detailed information concerning HpaR, please visit 2013 Peking iGEM Biosensor HpaR


Characterization

HpaR is of 17,235 Da (149 amino acids) that belongs to MarR family [4]. It performs as a repressor of the hpa cluster consisting of hpaGEDFHI genes (Fig.1), which participates in the catabolic pathway of 4-hydroxyphenylacetic acid (4HPAA) (Fig.2). HpaR derepress the downstream genes when contacting with ligands, including 4HPAA, 3-hydroxyphenylacetic acid (3HPAA) and 3, 4-dihydroxyphenylacetic acid (3,4-DHPAA).

Fig 1' The hpa cluster map in genome. The arrows inside the squares show the transcriptional direction of genes.Pr, Pg, Px, Pa and Pbc, represent the promoter controlling cognate clusters. Addition of 3-hydroxyphenylacetic acid can derepress the promoters inhibited by HpaR (for Pr and Pg activation) or HpaA (for Pbc activation). Finally the enzymes in these clusters will degrade the 3-HPAA to the intermediates in TCA cycle.

hpa cluster consists of three operons. The regulator gene, hpaR, is transcribed in the divert direction to other genes under Pr promoter. The adjacent promoter, Pg, initiates the transcription of the functional hpaGEDFHI operon. Pr and Pg are both regulated by HpaR and located in the intergenic region between the hpaR and hpaG (Fig.1).


Fig 2 The degradation pathway of 3HPAA. The letters on the arrows are the names of genes in the clusters. eg. HpaB,C means the step 1 is catalyzed by the product of hpaB and hpaC.


Sequence and Features

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 209
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Construction of biosensor circuit

Pg/HpaR biosensor circuit is constructed. The coding sequence of HpaR was obtained via PCR from E.coli.(Fig 3).

Fig 3' Schematic diagram of HpaR biosensor circuit. Orange arrowheads represent promoters. RBS is shown as green oval and red square stands for B0015 terminator.