Difference between revisions of "Part:BBa K2992019"

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===Usage and Biology===
 
===Usage and Biology===
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BgaR is the transcriptional regulator associated with the BgaR-BgaL lactose inducible system found naturally on the genome of <i>C. perfringens</i>. Our 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 <i>bgaR</i> as part of the entire regulatory unit comprising <i>bgaR</i>, and its divergent promoter P<i>bgaR</i> -P<i>bgaL</i>  with their associated 5’-UTR and RBS regions (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. <rb><br>
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BgaR is the transcriptional regulator associated with the BgaR-BgaL lactose inducible system (Plac) found naturally on the genome of <i>C. perfringens</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]).
  
 
===Characterisation===
 
===Characterisation===
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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.  See our [https://2019.igem.org/Team:Nottingham/Results results page] for more information.  
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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]).
  
 
https://static.igem.org/mediawiki/parts/e/e3/Plac_diagram.png
 
https://static.igem.org/mediawiki/parts/e/e3/Plac_diagram.png

Latest revision as of 03:54, 22 October 2019


bgaR gene from C. perfringens

Transcriptional regulator bgaR associated with a lactose inducible system from C. perfringens


Usage and Biology

BgaR is the transcriptional regulator associated with the BgaR-BgaL lactose inducible system (Plac) found naturally on the genome of C. perfringens. 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).

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
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 368
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 368
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 368
    Illegal BglII site found at 428
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 368
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 368
  • 1000
    COMPATIBLE WITH RFC[1000]

References

Cañadas, I., Groothuis, D., Zygouropoulou, M., Rodrigues, R. and Minton, N. (2019). RiboCas: A Universal CRISPR-Based Editing Tool for Clostridium. ACS Synthetic Biology, 8(6), pp.1379-1390.

Minton, N., Ehsaan, M., Humphreys, C., Little, G., Baker, J., Henstra, A., Liew, F., Kelly, M., Sheng, L., Schwarz, K. and Zhang, Y. (2016). A roadmap for gene system development in Clostridium. Anaerobe, 41, pp.104-112. 2019

Hartman, A., Liu, H. and Melville, S. (2010). Construction and Characterization of a Lactose-Inducible Promoter System for Controlled Gene Expression inClostridium perfringens. Applied and Environmental Microbiology, 77(2), pp.471-478.