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 being 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 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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<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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Revision as of 15:42, 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 genes are 2731bp, 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 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.