Difference between revisions of "Part:BBa K1590003"
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<partinfo>BBa_K1590003 short</partinfo>
 
<partinfo>BBa_K1590003 short</partinfo>
  
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The promoter pChr is suspected to be inducible by chromate.
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This promoter is found upstream of the <i>ChrBACF</i> - operon in <i>Ochrobactrum tritici</i> 5bvl1, located in the transposable element Tn<i>OtChr</i> of 7189bp length. P<i>chr</i> is suspected to be inducible by chromate via the chromate-responsive repressor ChrB.
  
  
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===Usage and Biology===
 
===Usage and Biology===
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This promoter is found upstream of the <i>ChrBACF</i> - operon in <i>Ochrobactrum tritici</i> 5bvl1, located in the transposable element Tn<i>OtChr</i> of 7189bp length.
 
  
 
Ochrobactrum tritici 5bvl1 was discovered in 2004 in a chromate contaminated wastewater treatment facility. A specific characteristic of this strain was that it could both, reduce Cr(VI) and be resistant to it at the same time. Previously characterised strains were either resistant or could reduce chromate. Over the last 10 years the group that discovered it started working out the underlying mechanisms of this set of characteristics.
 
Ochrobactrum tritici 5bvl1 was discovered in 2004 in a chromate contaminated wastewater treatment facility. A specific characteristic of this strain was that it could both, reduce Cr(VI) and be resistant to it at the same time. Previously characterised strains were either resistant or could reduce chromate. Over the last 10 years the group that discovered it started working out the underlying mechanisms of this set of characteristics.
  
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The <i>ChrBACF</i> - operon, expressed from the P<i>chr</i> - promoter led to the expression of the downstream genes <i>chrA, chrB, chrC</i>, and <i>chrF</i>. <i>chrA</i> was found to express and efflux pump, whereas the gene product of <i>chrB</i> was identified as being a chromate-inducible regulator of P<i>chr</i>. Additionally, <i>chrC</i> and <i>chrF</i> were found to confer tolerance to superoxides.
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The <i>ChrBACF</i> - operon, expressed from the P<i>chr</i> - promoter led to the expression of the downstream genes <i>chrA, chrB, chrC</i>, and <i>chrF</i>. <i>chrA</i> was found to express and efflux pump, whereas the gene product of <i>chrB</i> was identified as being a chromate-inducible regulator of P<i>chr</i>. Additionally, <i>chrC</i> and <i>chrF</i> were found to confer tolerance to superoxides, which can be byproducts of the reduction of Cr(VI).
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P<i>chr</i> itself is believed to be regulated by the chromate-sensitive repressor <i>chrB</i>. This has made it a candidate for a biological sensor of chromate contamination in e.g. rivers (Branco et al.), or in milk (iGEM BIT 2013). Our project focused on hijacking this circuit in order to build a detector for stainless steels.
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In our reporter construct, P<i>chr</i> was cloned upstream of <i>gfp</i>. The construct was fully sequence confirmed and confirmed in size.
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https://static.igem.org/mediawiki/2015/2/28/Dundee15_chr_pchrgfpinfo.png
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<br>
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Figure 1: Gel for size confirmation of pSB1C3-P<i>chr-gfp</i>. pSB1C3-P<i>chr-gfp</i> (3) was found to contain the expected sequence.
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 +
 
  
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P<i>chr itself is believed to be regulated by the chromate-sensitive repressor <i>chrB</i>. This has made it a candidate for a biological sensor of chromate contamination in e.g. rivers (Branco et al.), or in milk (iGEM BIT 2013).
 
  
  
 
===Functional Parameters===
 
===Functional Parameters===
 
<partinfo>BBa_K1590003 parameters</partinfo>
 
<partinfo>BBa_K1590003 parameters</partinfo>

Revision as of 17:37, 18 September 2015

Chromate responsive promoter


This promoter is found upstream of the ChrBACF - operon in Ochrobactrum tritici 5bvl1, located in the transposable element TnOtChr of 7189bp length. Pchr is suspected to be inducible by chromate via the chromate-responsive repressor ChrB.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 135
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Usage and Biology

Ochrobactrum tritici 5bvl1 was discovered in 2004 in a chromate contaminated wastewater treatment facility. A specific characteristic of this strain was that it could both, reduce Cr(VI) and be resistant to it at the same time. Previously characterised strains were either resistant or could reduce chromate. Over the last 10 years the group that discovered it started working out the underlying mechanisms of this set of characteristics.

The ChrBACF - operon, expressed from the Pchr - promoter led to the expression of the downstream genes chrA, chrB, chrC, and chrF. chrA was found to express and efflux pump, whereas the gene product of chrB was identified as being a chromate-inducible regulator of Pchr. Additionally, chrC and chrF were found to confer tolerance to superoxides, which can be byproducts of the reduction of Cr(VI).

Pchr itself is believed to be regulated by the chromate-sensitive repressor chrB. This has made it a candidate for a biological sensor of chromate contamination in e.g. rivers (Branco et al.), or in milk (iGEM BIT 2013). Our project focused on hijacking this circuit in order to build a detector for stainless steels.


In our reporter construct, Pchr was cloned upstream of gfp. The construct was fully sequence confirmed and confirmed in size.

Dundee15_chr_pchrgfpinfo.png
Figure 1: Gel for size confirmation of pSB1C3-Pchr-gfp. pSB1C3-Pchr-gfp (3) was found to contain the expected sequence.



Functional Parameters