Difference between revisions of "Part:BBa K2992021"
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===Usage===
 
===Usage===
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The BgaR-BgaL system of <i>C. perfringens</i>  comprises the transcriptional regulator BgaR belonging to the AraC-family and the β-galactosidase BgaL which are transcribed in a regulated fashion from the divergent P<i>gaR</i> -P<i>gaL</i>  promoter. The BgaR-BgaL system regulates the expression of carbohydrate metabolic genes in response to lactose concentrations (Hartman and Melville 2011). This parts entry represents the transcriptional terminator found downstream of the stop codon for bgaR P<i>gaR</i> -P<i>gaL</i> group has recently utlised this regulatory system in order to generate a tightly regulate inducible system for CRISPR-Cas mutagenesis in the genus <i>Clostridium</i> (Cañadas et al., 2019). In our project, we use the BgaR-BgaL  regulatory system (hyperlinks and descriptions) to drive the expression of our volatile and FAST reporter genes in an inducible fashion. Doing so helps us fulfil our goal of generating reporter strains for the prediction of botulinum neurotoxin production following food manufacture. The secondary structure for T<i>bgaR</i> was predicted using the Mfold web server (Zuker, 2003).  
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The BgaR-BgaL system of <i>C. perfringens</i>  comprises the transcriptional regulator BgaR belonging to the AraC-family and the β-galactosidase BgaL which are transcribed in a regulated fashion from the divergent P<i>gaR</i> -P<i>gaL</i>  promoter. The BgaR-BgaL system regulates the expression of carbohydrate metabolic genes in response to lactose concentrations (Hartman and Melville 2011). This part entry represents the transcriptional terminator found downstream of the stop codon for <i>bgaR</i>. The Plac system is comprised of the divergent P<i>bgaR</i>-P<i>bgaL</i> promoter and associated 5’-UTRs in conjunction with their cognate transcriptional regulator <i>bgaR</i>. The Plac system was used to drive lactose-dependent expression of <i>botR</i> ([https://parts.igem.org/Part:BBa_K2992002 BBa_K2992002]), which in turn, regulates the production of our reporter genes which we have placed under the control of a BotR-activated promoter ([https://parts.igem.org/Part:BBa_K2992028 BBa_K2992028], [https://parts.igem.org/Part:BBa_K2992029 BBa_K2992029], [https://parts.igem.org/Part:BBa_K2992036 BBa_K2992036]).
  
 
__NOTOC__
 
__NOTOC__
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[[Image:Mfold-K2992021-1.png]]
 
[[Image:Mfold-K2992021-1.png]]
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===Characterisation===
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The Plac system is an inducible promoter system utilizing the lactose operon from <i>C. perfringens</i>. This allowed us to test the maximum and minimum reporter expression range, informing the design of our electornic nose.  See our [https://2019.igem.org/Team:Nottingham/Results results page] for more information. In our project, the Plac system was used to drive lactose-dependent expression of <i>botR</i>  ([https://parts.igem.org/Part:BBa_K2992002 BBa_K2992002]) which in turn, regulates the production of our reporter genes which we have placed under the control of a BotR-activated promoter ([https://parts.igem.org/Part:BBa_K2992028 BBa_K2992028], [https://parts.igem.org/Part:BBa_K2992029 BBa_K2992029], [https://parts.igem.org/Part:BBa_K2992036 BBa_K2992036]).
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https://static.igem.org/mediawiki/parts/e/e3/Plac_diagram.png
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K2992021 SequenceAndFeatures</partinfo>
  
 
===References===
 
===References===
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Zuker 2003 update
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Zuker, M. (2003). Mfold web server for nucleic acid folding and hybridization prediction. Nucleic Acids Research, 31(13), pp.3406-3415.
  
 
<hr>
 
<hr>
 
'''Measurement'''
 
'''Measurement'''
 
* [http://openwetware.org/wiki/Cconboy:Terminator_Characterization/Results How these parts were measured]
 
* [http://openwetware.org/wiki/Cconboy:Terminator_Characterization/Results How these parts were measured]

Latest revision as of 03:55, 22 October 2019

Usage

The BgaR-BgaL system of C. perfringens comprises the transcriptional regulator BgaR belonging to the AraC-family and the β-galactosidase BgaL which are transcribed in a regulated fashion from the divergent PgaR -PgaL promoter. The BgaR-BgaL system regulates the expression of carbohydrate metabolic genes in response to lactose concentrations (Hartman and Melville 2011). This part entry represents the transcriptional terminator found downstream of the stop codon for bgaR. The Plac system is comprised of the divergent PbgaR-PbgaL promoter and associated 5’-UTRs in conjunction with their cognate transcriptional regulator bgaR. The Plac system was used to drive lactose-dependent expression of botR (BBa_K2992002), which in turn, regulates the production of our reporter genes which we have placed under the control of a BotR-activated promoter (BBa_K2992028, BBa_K2992029, BBa_K2992036).


Terminator.png

TbgaR transcriptional terminator from C. perfringens

  • TbgaR transcriptional terminator from C. perfringens

Secondary Structure

Mfold-K2992021-1.png

Characterisation

The Plac system is an inducible promoter system utilizing the lactose operon from C. perfringens. This allowed us to test the maximum and minimum reporter expression range, informing the design of our electornic nose. See our results page for more information. In our project, the Plac system was used to drive lactose-dependent expression of botR (BBa_K2992002) which in turn, regulates the production of our reporter genes which we have placed under the control of a BotR-activated promoter (BBa_K2992028, BBa_K2992029, BBa_K2992036).

Plac_diagram.png


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]

References

Zuker, M. (2003). Mfold web server for nucleic acid folding and hybridization prediction. Nucleic Acids Research, 31(13), pp.3406-3415.

Measurement

  • [http://openwetware.org/wiki/Cconboy:Terminator_Characterization/Results How these parts were measured]