Difference between revisions of "Team:UBonn HBRS/Description/Cloning"
(Created page with "{{UBonn_HBRS/header|title=Cloning|Project=active}} Content... {{UBonn_HBRS/footer}}") |
|||
| Line 1: | Line 1: | ||
{{UBonn_HBRS/header|title=Cloning|Project=active}} | {{UBonn_HBRS/header|title=Cloning|Project=active}} | ||
| − | + | == 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. | ||
| + | |||
| + | |||
| + | {{UBonn_HBRS/table-start|class=with-caption}} | ||
| + | <html> | ||
| + | <thead> | ||
| + | <tr> | ||
| + | <td>Gene name</td> | ||
| + | <td>Enzyme coded</td> | ||
| + | <td>Part number</td> | ||
| + | <td>Note</td> | ||
| + | </tr> | ||
| + | </thead> | ||
| + | <tbody> | ||
| + | <tr> | ||
| + | <td>XynA</td> | ||
| + | <td>Xylanase</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> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td>CelA</td> | ||
| + | <td>Cellulase</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> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td>EngB</td> | ||
| + | <td>Endoglucanase</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> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td>BsCel5c</td> | ||
| + | <td>Cellulase</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> | ||
| + | </tr> | ||
| + | </tbody> | ||
| + | </html> | ||
| + | {{UBonn_HBRS/table-end}} | ||
| + | <html><div class="caption"></html> | ||
| + | '''Table 1. Genes and Enzymes of Cellulase group.''' | ||
| + | <html></div></html> | ||
{{UBonn_HBRS/footer}} | {{UBonn_HBRS/footer}} | ||
Revision as of 23:45, 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 |
