Coding

Part:BBa_K2992019

Designed by: Yaseen Tengur   Group: iGEM19_Nottingham   (2019-09-16)
Revision as of 20:27, 21 October 2019 by AliceHodson (Talk | contribs) (→‎Characterisation)


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 found naturally on the genome of C. perfringens. Our group has recently utlised this regulatory system in order to generate a tightly regulate inducible system for CRISPR-Cas mutagenesis in the genus Clostridium (Cañadas et al., 2019). In our project, we use bgaR as part of the entire regulatory unit comprising bgaR, and its divergent promoter PbgaR -PbgaL 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.

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. See our results page for more information.

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.

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