Coding

Part:BBa_K2992019

Designed by: Yaseen Tengur   Group: iGEM19_Nottingham   (2019-09-16)
Revision as of 03:49, 22 October 2019 by JacobGausden (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. 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 botR (BBa_K2992002) in the genome of C. sporogenes. This, in turn, leads to expression of our chosen reporter genes in an inducible fashion.

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.

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