Difference between revisions of "Part:BBa K1682018:Experience"

(→‎Background - Lambert_GA 2020)
(→‎Background - Lambert_GA 2020)
Line 6: Line 6:
 
===Background - Lambert_GA 2020===
 
===Background - Lambert_GA 2020===
  
−
<br>
+
[[Image:Phomodelin.png|thumb|center|500px|<i>Figure 1: Diagram of the Pho Regulon signaling pathway. The Pho Regulon responds to extracellular P<sub>i</sub> levels and transcribes its regulatory genes.</i>]]
−
<center style="max-width: 700px; margin-left: auto;margin-right: auto;">
+
−
<img src="https://2020.igem.org/wiki/images/c/cf/T--Lambert_GA--NarBackground.jpg" style="width: 100%; background-color: #B8C9A2; padding: 20px; border-radius: 20px 20px 0px 0px;">
+
−
<p style="width: 100%; "><i style="width: 100%; text-align: center; display: block; background-color: #B8C9A2; border-radius: 0px 0px 20px 20px;">Figure 1. The diagram above shows the native Nar membrane-bound sensor proteins and the corresponding DNA-binding response regulators in <I>E. coli</I>.<br></br></i>
+
−
</p>
+
−
</center>
+
−
<br>
+
−
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.
+
  
 +
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.
+
===Experience - Lambert_GA 2020===
−
Nar Operon Background
+
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, [BBa_B1002 BBa_B1002], after super folder GFP and a rrnBT1 T1 terminator, [BBa_J61048 BBa_J61048], after TetR.
−
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===
 
===Applications of BBa_K1682018===

Revision as of 23:03, 27 October 2020


This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Background - Lambert_GA 2020

Figure 1: Diagram of the Pho Regulon signaling pathway. The Pho Regulon responds to extracellular Pi levels and transcribes its regulatory genes.

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.

Experience - Lambert_GA 2020

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, [BBa_B1002 BBa_B1002], after super folder GFP and a rrnBT1 T1 terminator, [BBa_J61048 BBa_J61048], after TetR.

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"

UNIQ884fbd68cd55ba20-partinfo-00000000-QINU UNIQ884fbd68cd55ba20-partinfo-00000001-QINU