Difference between revisions of "Part:BBa K1587004"
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<partinfo>BBa_K1587004 short</partinfo> | <partinfo>BBa_K1587004 short</partinfo> | ||
− | Constitutive promoter p(Bla). Crotonyl-coA reductase ccr coding region. 3-hydroxybutyryl-CoA dehydrogenase hbd coding region. 3-hydroxybutyryl-CoA dehydratase crt coding region. Acyl-CoA thioesterase 2 tesB coding region. acetyl-CoA acetyltransferase atoB coding region. Every genes are framed by strong RBS based on Ron Weiss thesis. This operon ends with an artificial terminator according to BBa_B1006. | + | ===BioBrick Description=== |
− | Due to a lack of time we decided to have this construction synthesised instead of using the assembly protocol. | + | |
+ | Constitutive promoter p(Bla). Crotonyl-coA reductase ccr coding region. 3-hydroxybutyryl-CoA dehydrogenase hbd coding region. 3-hydroxybutyryl-CoA dehydratase crt coding region. Acyl-CoA thioesterase 2 tesB coding region. acetyl-CoA acetyltransferase atoB coding region. Every genes are framed by strong RBS based on Ron Weiss thesis. This operon ends with an artificial terminator according to <a target="blank" href="https://parts.igem.org/Part:BBa_B1006">BBa_B1006</a>.<br> | ||
+ | Due to a lack of time we decided to have this construction synthesised instead of using the assembly protocol. | ||
+ | |||
+ | ===Usage and Biology=== | ||
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<center>[[File:TLSE SHLAGAGA.png]]</center> | <center>[[File:TLSE SHLAGAGA.png]]</center> | ||
<center>'''Figure 1: Genetic construct to produce butyrate via glucose'''</center> | <center>'''Figure 1: Genetic construct to produce butyrate via glucose'''</center> | ||
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<center>'''Figure 2: Metabolic pathway to produce butyrate via glucose using BBa_K1587004 in ''E. coli'''''</center> | <center>'''Figure 2: Metabolic pathway to produce butyrate via glucose using BBa_K1587004 in ''E. coli'''''</center> | ||
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
<partinfo>BBa_K1587004 SequenceAndFeatures</partinfo> | <partinfo>BBa_K1587004 SequenceAndFeatures</partinfo> | ||
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+ | Sequencing has been made thanks to 7 primers specifics for these genes: <br> | ||
+ | VF2, CCR, hbd, crt, testB, atoB and VR | ||
+ | |||
+ | |||
+ | ===Experiments=== | ||
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<center>[[File:TLSE Butprod.jpg]]</center> | <center>[[File:TLSE Butprod.jpg]]</center> | ||
− | '''Figure | + | '''Figure 3: NMR analysis of butyrate production in E. coli with BBa_K1587004 and wt''' |
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+ | Unfortunatly NMR analysis did not yield a significant over production of butyrate in E. coli. However we observed a statistically significant change in the production of other fermentation products such as ethanol or acetate. | ||
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Revision as of 16:30, 18 September 2015
Butyrate synthesis pathway regulated by constitutive promoter p(Bla)
BioBrick Description
Constitutive promoter p(Bla). Crotonyl-coA reductase ccr coding region. 3-hydroxybutyryl-CoA dehydrogenase hbd coding region. 3-hydroxybutyryl-CoA dehydratase crt coding region. Acyl-CoA thioesterase 2 tesB coding region. acetyl-CoA acetyltransferase atoB coding region. Every genes are framed by strong RBS based on Ron Weiss thesis. This operon ends with an artificial terminator according to <a target="blank" href="https://parts.igem.org/Part:BBa_B1006">BBa_B1006</a>.
Due to a lack of time we decided to have this construction synthesised instead of using the assembly protocol.
Usage and Biology
![TLSE SHLAGAGA.png](/wiki/images/c/c0/TLSE_SHLAGAGA.png)
![TLSE ButMetabo.jpg](/wiki/images/7/7d/TLSE_ButMetabo.jpg)
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 36
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1717
Illegal BamHI site found at 3634
Illegal XhoI site found at 1407 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 332
Illegal AgeI site found at 1254
Illegal AgeI site found at 2900 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1752
Illegal BsaI.rc site found at 2169
Illegal BsaI.rc site found at 3237
Illegal SapI site found at 123
Illegal SapI.rc site found at 3249
Sequencing has been made thanks to 7 primers specifics for these genes:
VF2, CCR, hbd, crt, testB, atoB and VR
Experiments
We used BBa_K1587004 in order to produce butyrate
![TLSE Butprod.jpg](/wiki/images/9/99/TLSE_Butprod.jpg)
Figure 3: NMR analysis of butyrate production in E. coli with BBa_K1587004 and wt
Unfortunatly NMR analysis did not yield a significant over production of butyrate in E. coli. However we observed a statistically significant change in the production of other fermentation products such as ethanol or acetate.