Difference between revisions of "Part:BBa K2317005"

 
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This part was designed to detect and degrade chlorophenol and control the population of the engineered bacteria at the mean time.  
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This part was designed to detect and degrade chlorophenol and control the population of the engineered bacteria at the mean time. <br>
  
 
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<partinfo>BBa_K2317005 parameters</partinfo>
 
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<head></head>
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<h3>Description</h3>
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<p>The function of the composite part we have been working on is to detect and degrade chlorophenol under population control of TA system.</p><br/>
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<div class="pic_box center">
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<img src="https://static.igem.org/mediawiki/2017/4/4d/T--Jilin_China--composite_parts01-01.png" width=”70%” /><br />
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Figure 1: The TA - DmpR - TfdB-JLU construct
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</div><br/>
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<p> This part is a combination of our Geneguard system and chlorophenol response system. Part BBa_J23107 is a constitutive promoter which could provide a constitutive expression of sensor DmpR and toxin CbtA. </p><br/>
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<p> This DmpR in our device is a mutant type based on the wild type DmpR used by 2013 Peking(<a href="https://parts.igem.org/Part:BBa_K1031211">BBa_K1031211</a>) and showed more efficient and wider substrate range than wild type.</p>
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<p> Toxin CbtA could keep the growth of the engineered bacterium in a low level. When exposed to contaminants, complex of DmpR and chlorophenol could bind to promoter Po and initiate the expression of antitoxin CbeA as well as TfdB-JLU, a monooxygenase that is responsible for initial hydroxylation of the benzene ring[1-2]. </p><br/>
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<div class="pic_box center">
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<img src="https://static.igem.org/mediawiki/2017/9/9a/T--Jilin_China--composite_parts05.png" width="60%" /><br />
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Figure 2. Reaction of TfdB-JLU
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</div><br/>
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<h3>Reference</h3>
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<p>[1] Ledger T, Pieper DH, Gonzalez B. (2006) Chlorophenol hydroxylases encoded by plasmid pJP4 differentially contribute to chlorophenoxyacetic acid degradation. Appl Environ Microbiol 72:2783–2792.</p><br/>
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<p>[2] Yang Lu. (2011) Cloning and characterisation of a novel 2,4-dichlorophenol hydroxylase from a metagenomic library derived from polychlorinated biphenyl-contaminated soil. Biotechnol Lett. 33:1159–1167</p><br/>
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Revision as of 15:45, 1 November 2017

TA - DmpR - TfdB-JLU

This part was designed to detect and degrade chlorophenol and control the population of the engineered bacteria at the mean time.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 147
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 625
    Illegal NgoMIV site found at 3116
    Illegal NgoMIV site found at 3779
    Illegal NgoMIV site found at 3805
    Illegal NgoMIV site found at 3890
    Illegal NgoMIV site found at 4430
    Illegal NgoMIV site found at 4621
    Illegal AgeI site found at 2572
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 310
    Illegal BsaI.rc site found at 851
    Illegal SapI.rc site found at 1508


Description

The function of the composite part we have been working on is to detect and degrade chlorophenol under population control of TA system.



Figure 1: The TA - DmpR - TfdB-JLU construct

This part is a combination of our Geneguard system and chlorophenol response system. Part BBa_J23107 is a constitutive promoter which could provide a constitutive expression of sensor DmpR and toxin CbtA.


This DmpR in our device is a mutant type based on the wild type DmpR used by 2013 Peking(BBa_K1031211) and showed more efficient and wider substrate range than wild type.

Toxin CbtA could keep the growth of the engineered bacterium in a low level. When exposed to contaminants, complex of DmpR and chlorophenol could bind to promoter Po and initiate the expression of antitoxin CbeA as well as TfdB-JLU, a monooxygenase that is responsible for initial hydroxylation of the benzene ring[1-2].



Figure 2. Reaction of TfdB-JLU

Reference

[1] Ledger T, Pieper DH, Gonzalez B. (2006) Chlorophenol hydroxylases encoded by plasmid pJP4 differentially contribute to chlorophenoxyacetic acid degradation. Appl Environ Microbiol 72:2783–2792.


[2] Yang Lu. (2011) Cloning and characterisation of a novel 2,4-dichlorophenol hydroxylase from a metagenomic library derived from polychlorinated biphenyl-contaminated soil. Biotechnol Lett. 33:1159–1167