Difference between revisions of "Part:BBa K2533033"

 
 
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It encodes glyceraldehyde-3-phosphate dehydrogenase.
 
It encodes glyceraldehyde-3-phosphate dehydrogenase.
  
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<h1>'''Usage and biology'''</h1>
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===Usage and Biology===
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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 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_K2533033 SequenceAndFeatures</partinfo>
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[[File:T--HUST-China--2018-tonglu-gapA.png ‎|400px|thumb|center|Figure1. RBS-gapA]]
  
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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-gel3.jpeg|400px|thumb|center|Figure2. Verification of successful transformation of pYYDT-gapA]]
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Our target gene is 1126bp, 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 gapA could be overexpressed by engineered Shewanella, we did Real-Time Quantitative PCR.
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[[File:T--HUST–China--2018-result-fig3.png ‎|400px|thumb|center|Figure3. Relative expression level of gapA in engineered Shewanella Oneidensis MR-1.]]
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There was no signal in bacteria which contained pYYDT so we chose pYYDT-gapA as standard quantity.
  
 
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===Functional Parameters===
 
===Functional Parameters===
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<partinfo>BBa_K2533033 parameters</partinfo>
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<partinfo>BBa_K2533030 parameters</partinfo>
 
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Latest revision as of 22:53, 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 to be 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 gene is 1126bp, 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 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.