Difference between revisions of "Part:BBa K2992024"

 
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<partinfo>BBa_K2992024 short</partinfo>
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5’UTR containing RBS predicted to regulate <i>bgaL</i> in <i>C. perfringens</i>.   
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===Usage and Biology===
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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 RBS and 5’-UTR predicted to regulate <i>bgaL</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]).
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===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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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_K2992023 SequenceAndFeatures</partinfo>
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===References===
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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
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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 RiboCas - update
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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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Zuker, M. (2003). Mfold web server for nucleic acid folding and hybridization prediction. Nucleic Acids Research, 31(13), pp.3406-3415.
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<!-- Uncomment this to enable Functional Parameter display
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===Functional Parameters===
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<partinfo>BBa_K2992024 parameters</partinfo>
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Latest revision as of 03:57, 22 October 2019


5-UTR containing RBS for bgaL from C. perfringens

5’UTR containing RBS predicted to regulate bgaL in C. perfringens.

Usage and Biology

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 parts entry represents the RBS and 5’-UTR predicted to regulate bgaL. 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. See our results page for more information.

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

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 RiboCas - update

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.


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