Difference between revisions of "Part:BBa K2533033"
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<h1>'''Characterization'''</h1> | <h1>'''Characterization'''</h1> | ||
This is one section for NADH production part. | This is one section for NADH production part. | ||
| − | [[File:T--HUST-China--2018-tonglu-gapA.png |400px|thumb|center|Figure1 | + | [[File:T--HUST-China--2018-tonglu-gapA.png |400px|thumb|center|Figure1. RBS-gapA]] |
<h2>DNA Gel Electrophoretic</h2> | <h2>DNA Gel Electrophoretic</h2> | ||
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-gel3.jpeg|400px|thumb|center|Figure2 | + | [[File:T--HUST-China--2018-Notebook-gel3.jpeg|400px|thumb|center|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. | 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. | ||
<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.png |400px|thumb|center|Figure3 | + | [[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. | ||
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.
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.
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.
There was no signal in bacteria which contained pYYDT so we chose pYYDT-gapA as standard quantity.