Difference between revisions of "Team:UBonn HBRS/Description/Cloning"
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| Line 33: | Line 33: | ||
<td>Xylanase</td> | <td>Xylanase</td> | ||
<td>BBa_K2029002</td> | <td>BBa_K2029002</td> | ||
| − | <td><a href="http://www.microbialcellfactories.com/content/11/1/66">http://www.microbialcellfactories.com/content/11/1/66</a></td> | + | <td><a href="http://www.microbialcellfactories.com/content/11/1/66" target="_blank">http://www.microbialcellfactories.com/content/11/1/66</a></td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
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<td>Cellulase</td> | <td>Cellulase</td> | ||
<td>BBa_K2029004</td> | <td>BBa_K2029004</td> | ||
| − | <td><a href="https://www.ncbi.nlm.nih.gov/pubmed/22101447">https://www.ncbi.nlm.nih.gov/pubmed/22101447</a></td> | + | <td><a href="https://www.ncbi.nlm.nih.gov/pubmed/22101447" target="_blank">https://www.ncbi.nlm.nih.gov/pubmed/22101447</a></td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
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<td>Endoglucanase</td> | <td>Endoglucanase</td> | ||
<td>BBa_K2029005</td> | <td>BBa_K2029005</td> | ||
| − | <td><a href="http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1785254/">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1785254/</a></td> | + | <td><a href="http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1785254/" target="_blank">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1785254/</a></td> |
</tr> | </tr> | ||
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<td>Cellulase</td> | <td>Cellulase</td> | ||
<td>BBa_K2029006</td> | <td>BBa_K2029006</td> | ||
| − | <td><a href="http://www.ncbi.nlm.nih.gov/pubmed/21549854">http://www.ncbi.nlm.nih.gov/pubmed/21549854</a></td> | + | <td><a href="http://www.ncbi.nlm.nih.gov/pubmed/21549854" target="_blank">http://www.ncbi.nlm.nih.gov/pubmed/21549854</a></td> |
</tr> | </tr> | ||
</tbody> | </tbody> | ||
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'''Table 1. Genes and Enzymes of Cellulase group.''' | '''Table 1. Genes and Enzymes of Cellulase group.''' | ||
<html></div></html> | <html></div></html> | ||
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| + | |||
| + | === Esterase-Laccase group === | ||
| + | Enzymes which hydrolyse esters or oxidise phenols and aromatic compounds found in commercial inks. Their ability to attack ink particles is not well studied. | ||
{{UBonn_HBRS/footer}} | {{UBonn_HBRS/footer}} | ||
Revision as of 23:48, 19 October 2016
Cloning
General Abstract
The cloning aspect of this project is responsible for programming bacterial cells to secrete enzymes which can decouple ink particles from paper fibers.
To this end, genes coding for enzymes were synthesised and ligated into a plasmid backbone, which is designed for use in both Escherichia coli and Bacillus subtilis.
As expression of enzymes is intended for both Bacillus subtilis and E.coli, there are two separate tracks for cloning. For B. subtilis, the genes were assembled into pSBbs1AK1 along with secretory tags, and for E.coli, an inductive promoter (pLac) is placed upstream of the genes.
Genes of Interest
Within the scope of this project, genes of interest (GOIs) are those which catalyse either cellulolysis of paper fiber, or hydrolysis of ink particles.
Synthesised at GenScript, each of these genes includes a sequence coding for His-tag, for ease of protein purification.
Cellulase group
Enzymes which break down cellulose molecules into monosaccharides, therefore partially disintegrate paper fibers and set ink particles free into the supernatant.
| Gene name | Enzyme coded | Part number | Note |
| XynA | Xylanase | BBa_K2029002 | http://www.microbialcellfactories.com/content/11/1/66 |
| CelA | Cellulase | BBa_K2029004 | https://www.ncbi.nlm.nih.gov/pubmed/22101447 |
| EngB | Endoglucanase | BBa_K2029005 | http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1785254/ |
| BsCel5c | Cellulase | BBa_K2029006 | http://www.ncbi.nlm.nih.gov/pubmed/21549854 |
Esterase-Laccase group
Enzymes which hydrolyse esters or oxidise phenols and aromatic compounds found in commercial inks. Their ability to attack ink particles is not well studied.
