Difference between revisions of "Part:BBa K2317005"
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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/> | <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/> | ||
<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> | <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> | ||
| − | <p> Toxin CbtA could keep the growth of the engineered | + | <p> Toxin CbtA could keep the growth of the engineered bacteria 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/> |
<div class="pic_box center"> | <div class="pic_box center"> | ||
<img src="https://static.igem.org/mediawiki/2017/9/9a/T--Jilin_China--composite_parts05.png" width="60%" /><br /> | <img src="https://static.igem.org/mediawiki/2017/9/9a/T--Jilin_China--composite_parts05.png" width="60%" /><br /> | ||
Revision as of 23:43, 1 November 2017
TA - DmpR - TfdB-JLU
This part was designed to detect and degrade chlorophenol and control the population of the engineered bacteria in the meantime.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 147
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE 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 - 1000INCOMPATIBLE 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 bacteria 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