Difference between revisions of "Part:BBa K2533033"

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<h2>Real-Time Quantitative PCR</h2>
 
<h2>Real-Time Quantitative PCR</h2>
 
To demonstrate that gapA could be overexpressed by engineered Shewanella, we did Real-Time Quantitative PCR.  
 
To demonstrate that gapA could be overexpressed by engineered Shewanella, we did Real-Time Quantitative PCR.  
[[File:T--HUST–China--2018-result-fig3.jpeg ‎|400px|thumb|center|Figure3:Relative expression level of gapA in engineered Shewanella Oneidensis MR-1.]]
+
[[File:T--HUST–China--2018-result-fig3.png ‎|400px|thumb|center|Figure3:Relative expression level of gapA in engineered Shewanella Oneidensis MR-1.]]
 
There was no signal in bacteria which contained pYYDT so we chose pYYDT-gapA as standard quantity.
 
There was no signal in bacteria which contained pYYDT so we chose pYYDT-gapA as standard quantity.
  

Revision as of 15:12, 17 October 2018


RBS-gapA

It encodes glyceraldehyde-3-phosphate dehydrogenase.

Usage and biology

It encodes glyceraldehyde-3-phosphate dehydrogenase which could transform 3- phosphoglyceraldehyde into 1,3- diphosphoglycerate. With the overexpression of gapA, Shewanella could produce NADH more efficiently, which brings more electricity being produced.

Characterization

This is one section for NADH production part.

Figure1:RBS-gapA

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-gapA

Our target genes are 1126bp, and as the marker is DS5000, we could be sure that the bright bands in this picture are our target genes.

Real-Time Quantitative PCR

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

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

There was no signal in bacteria which contained pYYDT so we chose pYYDT-gapA as standard quantity.