Device

Part:BBa_K1682018:Experience

Designed by: Nikola Dyulgyarov   Group: iGEM15_HKUST-Rice   (2015-09-17)
Revision as of 22:50, 27 October 2020 by Saifkhan (Talk | contribs) (→‎Background - Lambert_GA 2020)


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Background - Lambert_GA 2020

Background - Lambert_GA 2020

Native to E. coli, the Nar Operon regulates anaerobic gene expression in response to two electron acceptors: nitrate and nitrite. This system consists of two homologous membrane-bound sensor proteins (NarX and NarQ) as well as two homologous DNA-binding response regulators (NarL and NarP). NarL’s conjugate is the membrane-bound NarX protein while NarP’s conjugate is the membrane-bound NarQ protein. Lambert iGEM is utilizing this system to test nitrate and nitrite levels, NarL and NarP expression, and downstream GFP expression through mathematical models.


To optimize functionality, Lambert_GA altered the original part designed by HKUST-Rice 2015 by replacing the original terminators with rrNB T1 terminator and T7Te terminator after super folder GFP and a rrnBT1 T1 terminator after TetR. Nar Operon Background Native to E. coli, the Nar Operon regulates anaerobic gene expression in response to two electron acceptors: nitrate and nitrite. This system consists of two homologous membrane-bound sensor proteins (NarX and NarQ) as well as two homologous DNA-binding response regulators (NarL and NarP). NarL’s conjugate is the membrane-bound NarX protein while NarP’s conjugate is the membrane-bound NarQ protein. Our team is utilizing this system to test nitrate and nitrite levels, NarL and NarP expression, and downstream GFP expression through mathematical models.

Applications of BBa_K1682018

Lambert_GA 2020 is working on demonstrating that this part can be transformed into competent E.coli cells while retaining functionality. Attempts at ligation both with restriction digest cloning and Gibson Assembly cloning failed. Our team is continuing to troubleshoot and is still in the cloning process. In 2021, we plan on utilizing this to model a nitrate biosensor in hydroponic/aquaponic systems.

src="T--Lambert_GA--_Nardiagram.png"

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