Difference between revisions of "Part:BBa K2533053"
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<h2>DNA Gel Electrophoretic</h2> | <h2>DNA Gel Electrophoretic</h2> | ||
| − | To make sure that we get the target | + | To make sure that we get the target genes, we did DNA gel electrophoretic for verification. And here is the result. |
[[File:T--HUST-China--2018-jiaotu-mleS-lldP-ldhA.png|400px|thumb|center|Figure2:Verification of successful transformation of pSB1C3-RBS-mleS-RBS-lldP-RBS-ldhA-TT]] | [[File:T--HUST-China--2018-jiaotu-mleS-lldP-ldhA.png|400px|thumb|center|Figure2:Verification of successful transformation of pSB1C3-RBS-mleS-RBS-lldP-RBS-ldhA-TT]] | ||
Our target genes are 4388bp, 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 4388bp, and as the marker is DS5000, we could be sure that the bright bands in this picture are our target genes. | ||
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By detecting the production of lactate after expressing, we might find out whether RBS-mleS-RBS-lldP-RBS-ldhA-TT could effectively help Rhodopseudomonas palustris transport produced lactate out of the cell. | By detecting the production of lactate after expressing, we might find out whether RBS-mleS-RBS-lldP-RBS-ldhA-TT could effectively help Rhodopseudomonas palustris transport produced lactate out of the cell. | ||
[[File:T--HUST-China--2018-expression of lactate.png |400px|thumb|center|Figure3:shows that our modification is effective. Every gene circuits can help strains produce lactate, and mleS-lldP-ldhA is the most efficient one. Therefore, our construction of gene circuits achieves the goal to help strains produce lactate.]] | [[File:T--HUST-China--2018-expression of lactate.png |400px|thumb|center|Figure3:shows that our modification is effective. Every gene circuits can help strains produce lactate, and mleS-lldP-ldhA is the most efficient one. Therefore, our construction of gene circuits achieves the goal to help strains produce lactate.]] | ||
| − | It could be demonstrated that | + | It could be demonstrated that the target genes could be expressed in the engineered cells. More lactate has been produced and transported by engineered bacteria. |
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===Functional Parameters=== | ===Functional Parameters=== | ||
<partinfo>BBa_K2533030 parameters</partinfo> | <partinfo>BBa_K2533030 parameters</partinfo> | ||
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Revision as of 17:38, 17 October 2018
RBS-mleS-RBS-lldP-RBS-ldhA-TT
malate dehydrogenase & L-lactate permease & D-lactate dehydrogenase
Usage and biology
It encodes malate dehydrogenase, L-lactate permease and D-lactate dehydrogenase, which could produce and transport more lactate. mleS could convert malic acid to L-lactate, lldP could transport lactate out of the cell. ldhA could convert pyruvate to D-lactate. In this way, Rhodopseudomonas palustris could efficiently transport produced lactate out of the cell, which later will be used by Shewanella.
Characterization
This is one section for lactate production and transportation part.
DNA Gel Electrophoretic
To make sure that we get the target genes, we did DNA gel electrophoretic for verification. And here is the result.
Our target genes are 4388bp, and as the marker is DS5000, we could be sure that the bright bands in this picture are our target genes.
Electrogenesis
By detecting the production of lactate after expressing, we might find out whether RBS-mleS-RBS-lldP-RBS-ldhA-TT could effectively help Rhodopseudomonas palustris transport produced lactate out of the cell.
It could be demonstrated that the target genes could be expressed in the engineered cells. More lactate has been produced and transported by engineered bacteria.