Difference between revisions of "Part:BBa K2533036"

 
 
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It encodes pyruvate formate-lyase.
 
It encodes pyruvate formate-lyase.
  
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<h1>'''Usage and biology'''</h1>
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===Usage and Biology===
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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.
  
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<h1>'''Characterization'''</h1>
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<span class='h3bb'>Sequence and Features</span>
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This is one section for NADH production part.
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<partinfo>BBa_K2533036 SequenceAndFeatures</partinfo>
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[[File:T--HUST-China--2018-tonglu-pflB.png ‎|400px|thumb|center|Figure1. RBS-pflB]]
  
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<h2>DNA Gel Electrophoretic</h2>
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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.
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[[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 gene is 2731bp, and as the marker is DS5000, we could be sure that the bright band in this picture is our target gene.
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<h2>Real-Time Quantitative PCR</h2>
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To demonstrate that mdh could be overexpressed by engineered Shewanella, we did Real-Time Quantitative PCR.
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[[File:T--HUST–China--2018-result-pflB.jpeg ‎|400px|thumb|center|Figure3. Relative expression level of pflB in engineered Shewanella Oneidensis MR-1.]]
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As we can see from this figure, pflB could be overexpressed by engineered Shewanella.
  
 
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===Functional Parameters===
 
===Functional Parameters===
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<partinfo>BBa_K2533036 parameters</partinfo>
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<partinfo>BBa_K2533030 parameters</partinfo>
 
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Latest revision as of 22:56, 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.