Difference between revisions of "Part:BBa K2533036"
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To make sure that we get the target gene, we did the DNA gel electrophoretic to separate different gene. And here is the result.
 
To make sure that we get the target gene, we did the DNA gel electrophoretic to separate different gene. And here is the result.
 
[[File:T--HUST-China--2018-Notebook-gel1.jpeg|400px|thumb|center|Figure2:Verification of successful transformation of pYYDT-pflB]]
 
[[File:T--HUST-China--2018-Notebook-gel1.jpeg|400px|thumb|center|Figure2:Verification of successful transformation of pYYDT-pflB]]
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Our target genes are 2731bp, and as the marker is DS5000, we could be sure that the bright bands in this picture are our target genes.
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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.
  
 
<h2>Real-Time Quantitative PCR</h2>
 
<h2>Real-Time Quantitative PCR</h2>

Revision as of 16:55, 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 being 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.