Difference between revisions of "Part:BBa K2533034"
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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-mdh.png |400px|thumb|center|Figure1 | + | [[File:T--HUST-China--2018-tonglu-mdh.png |400px|thumb|center|Figure1. RBS-mdh]] |
<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-mdh]] |
Our target gene is 1145bp, 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 1145bp, 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-mdh.png |400px|thumb|center|Figure3 | + | [[File:T--HUST–China--2018-result-mdh.png |400px|thumb|center|Figure3. Relative expression level of mdh in engineered Shewanella Oneidensis MR-1.]] |
As we can see from this figure, mdh could be overexpressed by engineered Shewanella. | As we can see from this figure, mdh could be overexpressed by engineered Shewanella. | ||
Revision as of 22:54, 17 October 2018
RBS-mdh
It encodes NAD dependent malate dehydrogenase.
Usage and biology
It encodes NAD dependent malate dehydrogenase which transforms malate into pyruvate. With the overexpression of mdh, 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 1145bp, 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, mdh could be overexpressed by engineered Shewanella.