Difference between revisions of "Part:BBa K1465110"
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===Usage and Biology=== | ===Usage and Biology=== | ||
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| + | <h4>C4 Carboxylate Antiporter DcuB</h4> | ||
| + | Under anaerobic conditions <i>E. coli</i> cells use different alternative electron acceptors instead of oxygen. Partially the bacteria use fumarate respiration, whereby fumarate is reduced into succinate. There are also other potential less-oxidizing substances for bacteria to release their electrons, for example anorganic compounds like nitrate (NO<sub>3</sub><sup>-</sup>) or sulfate (SO<sub>4</sub><sup>2-</sup>).(<a href="#Gottschalk1986">Gottschalk <i>et al.</i>, 1986</a>) | ||
| + | Fumarate respiration leads to succinate excretion through the C4 carboxylate transporter DcuB. It is an antiporter which exchanges fumarate against succinate under anaerobic conditions. Under aerobic condition there is usually no succinate release observed. In connection to the carbon dioxide fixation in our second module we planned on working under oxygen limiting conditions, hence effective carbon dioxid fixation is possible. So in case of oxygen limiting conditions, there could occured partial fumarate respiration in <i>E. coli</i>. Besides there was shown activity of DcuB antiporter in the presence of high fumarate concentrations (<a href="#Janausch2001">Janausch, 2001</a>). To achieve an effective electron uptake and prevent any succinate excretion, the C4 carboxylate antiporter DcuB has to be knocked out in our <i>E. coli</i> strain. <br> | ||
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| + | We planned a targeted knockout of the <i>dcuB</i> gene in <i>E. coli</i> KRX using the <a href="http://www.genebridges.com/storage/Manuals_PDF/K006%20Ecoli%20Gene%20Deletion%20Kit-version2.3-2012.pdf">Genebridge Red/ET-System</a>. In the same step we are going to integrate the outer membrane porine OprF (<a href="https://parts.igem.org/Part:BBa_K1172507">BBa_K1172507</a>) into the bacterial chromosome under controll of a constitutive promotor (<a href="https://parts.igem.org/Part:BBa_J23104">BBa_J23104</a>). This ensure the permeability of the outer membrane and avoid a plasmid overload of the bacteria, because for our system the outer membrane porines are indispensable.<br> | ||
| + | </html> | ||
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Revision as of 18:36, 20 October 2014
dcuB rev
dcuB rev
Usage and Biology
Usage and Biology
C4 Carboxylate Antiporter DcuB
Under anaerobic conditions E. coli cells use different alternative electron acceptors instead of oxygen. Partially the bacteria use fumarate respiration, whereby fumarate is reduced into succinate. There are also other potential less-oxidizing substances for bacteria to release their electrons, for example anorganic compounds like nitrate (NO3-) or sulfate (SO42-).(Gottschalk et al., 1986) Fumarate respiration leads to succinate excretion through the C4 carboxylate transporter DcuB. It is an antiporter which exchanges fumarate against succinate under anaerobic conditions. Under aerobic condition there is usually no succinate release observed. In connection to the carbon dioxide fixation in our second module we planned on working under oxygen limiting conditions, hence effective carbon dioxid fixation is possible. So in case of oxygen limiting conditions, there could occured partial fumarate respiration in E. coli. Besides there was shown activity of DcuB antiporter in the presence of high fumarate concentrations (Janausch, 2001). To achieve an effective electron uptake and prevent any succinate excretion, the C4 carboxylate antiporter DcuB has to be knocked out in our E. coli strain.We planned a targeted knockout of the dcuB gene in E. coli KRX using the Genebridge Red/ET-System. In the same step we are going to integrate the outer membrane porine OprF (BBa_K1172507) into the bacterial chromosome under controll of a constitutive promotor (BBa_J23104). This ensure the permeability of the outer membrane and avoid a plasmid overload of the bacteria, because for our system the outer membrane porines are indispensable.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]