Difference between revisions of "Part:BBa K2533053"
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<partinfo>BBa_K2533053 short</partinfo>
 
<partinfo>BBa_K2533053 short</partinfo>
  
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Production and transport of lactic acid
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malate dehydrogenase & L-lactate permease & D-lactate dehydrogenase
  
 
<h1>'''Usage and biology'''</h1>
 
<h1>'''Usage and biology'''</h1>
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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.
+
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.
  
 
<h1>'''Characterization'''</h1>
 
<h1>'''Characterization'''</h1>
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This is one section for lactate utilization part.
+
This is one section for lactate production and transportation part.
 
[[File:T--HUST-China--2018-tonglu-mleS-lldP-ldhA.png ‎|400px|thumb|center|Figure1:RBS-mleS-RBS-lldP-RBS-ldhA-TT]]
 
[[File:T--HUST-China--2018-tonglu-mleS-lldP-ldhA.png ‎|400px|thumb|center|Figure1:RBS-mleS-RBS-lldP-RBS-ldhA-TT]]
  
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<h2>Electrogenesis</h2>
 
<h2>Electrogenesis</h2>
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By detecting the production of lactate after expressing protein, we might find out whether ldhA could effectively help Rhodopseudomonas palustris to produce more lactate.
+
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 targeted genes could be expressed in the engineered cells. More lactate has been produced by engineered bacteria.
+
It could be demonstrated that targeted genes could be expressed in the engineered cells. More lactate has been produced and transported 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 16:58, 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.

Figure1:RBS-mleS-RBS-lldP-RBS-ldhA-TT

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-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.

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.

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 targeted genes could be expressed in the engineered cells. More lactate has been produced and transported by engineered bacteria.