Difference between revisions of "Part:BBa K2533036"

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<h1>'''Usage and biology'''</h1>
 
<h1>'''Usage and biology'''</h1>
It encodes pyruvate formate-lyase to transform pyruvate into Acetyl-CoA. With the overexpression of pflB, Shewanella could produce NADH more efficiently, which brings more electricity being produced.
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It encodes pyruvate formate-lyase to transform pyruvate into Acetyl-CoA. With the overexpression of pflB, Shewanella could produce NADH more efficiently, which brings more electricity to be produced.
  
 
<h1>'''Characterization'''</h1>
 
<h1>'''Characterization'''</h1>

Revision as of 17:21, 17 October 2018


RBS-pflB

It encodes pyruvate formate-lyase.

Usage and biology

It encodes pyruvate formate-lyase to transform pyruvate into Acetyl-CoA. With the overexpression of pflB, Shewanella could produce NADH more efficiently, which brings more electricity to be produced.

Characterization

This is one section for NADH production part.

Figure1:RBS-pflB

DNA Gel Electrophoretic

To make sure that we get the target gene, we did the DNA gel electrophoretic to separate different gene. And here is the result.

Figure2:Verification of successful transformation of pYYDT-pflB

Our target gene is 2731bp, and as the marker is DS5000, we could be sure that the bright band in this picture is our target gene.

Real-Time Quantitative PCR

To demonstrate that mdh could be overexpressed by engineered Shewanella, we did Real-Time Quantitative PCR.

Figure3:Relative expression level of pflB in engineered Shewanella Oneidensis MR-1.

As we can see from this figure, pflB could be overexpressed by engineered Shewanella.