Difference between revisions of "Part:BBa K2533047"
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<h1>'''Usage and biology'''</h1>
 
<h1>'''Usage and biology'''</h1>
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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.
+
ldhA refers to fermentative D-lactate dehydrogenase, NAD-dependent, performing the conversion of pyruvate to D-lactate. In this way, Rhodopseudomonas palustris could produce lactate more efficiently, which brings Shewanella more carbon sources.
  
 
<h1>'''Characterization'''</h1>
 
<h1>'''Characterization'''</h1>
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<h2>DNA Gel Electrophoretic</h2>
 
<h2>DNA Gel Electrophoretic</h2>
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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 DNA gel electrophoretic for verification. And here is the result.
 
[[File:T--HUST-China--2018-jiaotu-ldha and lldp.png|400px|thumb|center|Figure2:Verification of successful transformation of pSB1C3-RBS-ldhA]]
 
[[File:T--HUST-China--2018-jiaotu-ldha and lldp.png|400px|thumb|center|Figure2:Verification of successful transformation of pSB1C3-RBS-ldhA]]
 
Our target genes are 1047bp, 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 1047bp, and as the marker is DS5000, we could be sure that the bright bands in this picture are our target genes.
  
 
<h2>Electrogenesis</h2>
 
<h2>Electrogenesis</h2>
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By comparing the ability of producing electricity, we might find out whether dld could effectively help Shewanella to produce more electricity.
+
By detecting the production of lactate after expressing protein, we might find out whether ldhA could effectively help Rhodopseudomonas palustris to produce more 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 achieve 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 achieve the goal to help strains produce lactate.]]
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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.  
+
It could be demonstrated that targeted genes could be expressed in the engineered cells. More lactate has been produced by engineered bacteria.
−
 
+
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  
 
===Functional Parameters===
 
===Functional Parameters===
 
<partinfo>BBa_K2533030 parameters</partinfo>
 
<partinfo>BBa_K2533030 parameters</partinfo>
 
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Revision as of 15:16, 17 October 2018


RBS-ldhA

Production of lactate

Usage and biology

ldhA refers to fermentative D-lactate dehydrogenase, NAD-dependent, performing the conversion of pyruvate to D-lactate. In this way, Rhodopseudomonas palustris could produce lactate more efficiently, which brings Shewanella more carbon sources.

Characterization

This is one section for lactate utilization part.

Figure1:RBS-ldhA

DNA Gel Electrophoretic

To make sure that we get the target gene, we did DNA gel electrophoretic for verification. And here is the result.

Figure2:Verification of successful transformation of pSB1C3-RBS-ldhA

Our target genes are 1047bp, 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 protein, we might find out whether ldhA could effectively help Rhodopseudomonas palustris to produce more lactate.

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 achieve the goal to help strains produce lactate.

It could be demonstrated that targeted genes could be expressed in the engineered cells. More lactate has been produced by engineered bacteria.