Difference between revisions of "Part:BBa K2533036"
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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-pflB.png |400px|thumb|center|Figure1 | + | [[File:T--HUST-China--2018-tonglu-pflB.png |400px|thumb|center|Figure1. RBS-pflB]] |
<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-gel1.jpeg|400px|thumb|center|Figure2 | + | [[File:T--HUST-China--2018-Notebook-gel1.jpeg|400px|thumb|center|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. | 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. | ||
<h2>Real-Time Quantitative PCR</h2> | <h2>Real-Time Quantitative PCR</h2> | ||
To demonstrate that mdh could be overexpressed by engineered Shewanella, we did Real-Time Quantitative PCR. | To demonstrate that mdh could be overexpressed by engineered Shewanella, we did Real-Time Quantitative PCR. | ||
| − | [[File:T--HUST–China--2018-result-pflB.jpeg |400px|thumb|center|Figure3 | + | [[File:T--HUST–China--2018-result-pflB.jpeg |400px|thumb|center|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. | As we can see from this figure, pflB could be overexpressed by engineered Shewanella. | ||
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.
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 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.
As we can see from this figure, pflB could be overexpressed by engineered Shewanella.