Difference between revisions of "Part:BBa K2533030"
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<h1>'''Characterization'''</h1> | <h1>'''Characterization'''</h1> | ||
This is one section for lactate utilization part. | This is one section for lactate utilization part. | ||
| − | [[File:T--HUST-China--2018- | + | [[File:T--HUST-China--2018-tonglu-dld.png |400px|thumb|center|Figure1:RBS-dld]] |
<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-gel9.jpeg|400px|thumb|center|Figure1:Verification of successful transformation of pYYDT-dld]] | |
Our target genes are 3366bp, and as the marker is DS5000, we could be sure that the bright bands in this picture are our target genes. | Our target genes are 3366bp, and as the marker is DS5000, we could be sure that the bright bands in this picture are our target genes. | ||
<h2>Real-Time Quantitative PCR</h2> | <h2>Real-Time Quantitative PCR</h2> | ||
To demonstrate that dld could be overexpressed by engineered Shewanella, we did Real-Time Quantitative PCR. | To demonstrate that dld could be overexpressed by engineered Shewanella, we did Real-Time Quantitative PCR. | ||
| − | + | [[File:T--HUST–China--2018-result-fig1.jpeg |400px|thumb|center|Figure1:Relative expression level of dld in engineered Shewanella Oneidensis MR-1.]] | |
As we can see from this figure, dld could be overexpressed by engineered Shewanella. For further verification, we used the engineered bacteria to produce electricity. | As we can see from this figure, dld could be overexpressed by engineered Shewanella. For further verification, we used the engineered bacteria to produce electricity. | ||
Revision as of 09:53, 17 October 2018
RBS-dld
It encodes D-lactate dehydrogenase.
Usage and biology
dld refers to FAD-dependent D-lactate dehydrogenase which could catalyze D-lactate’s transformation into pyruvate. With the overexpression of dld, Shewanella could utilize D-lactate more efficiently, which brings more electricity being produced.
Characterization
This is one section for lactate utilization 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 genes are 3366bp, 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 dld could be overexpressed by engineered Shewanella, we did Real-Time Quantitative PCR.
As we can see from this figure, dld could be overexpressed by engineered Shewanella. For further verification, we used the engineered bacteria to produce electricity.
Electrogenesis
By comparing the ability of producing electricity, we might find out whether dld could effectively help Shewanella to produce more electricity. tu It could be demonstrated that targeted genes could be expressed in the engineered cells. More NADH has been produced by engineered bacteria, which helps to produce more electricty.