Team:UGent/Results
From 2013.igem.org
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</p> | </p> | ||
<h1> UV Test </h1> | <h1> UV Test </h1> | ||
- | <p> Phage transduction was used to perform the deletion of the <i>recA</i> gene. To check whether the gene was successfully knocked out, an UV test was developed. The UV light puts a certain amount of stress on the bacterial cells. | + | <p> Phage transduction was used to perform the deletion of the <i>recA</i> gene after chromosomal evolution. To check whether the gene was successfully knocked out, an UV test was developed. The UV light puts a certain amount of stress on the bacterial cells. In respons to this stress, an SOS pathway is triggered in which <i>recA</i> plays an important role. Cells in which the <i>recA</i> gene is deleted will not survive on the UV-treated plate. Colonies in which <i>recA</i> was successfully deleted still can be found on a non-treated backup plate. |
<br><br> | <br><br> | ||
<b>RESULT: </b> | <b>RESULT: </b> | ||
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<li>15’’ – still some colonies on UV-part of the plate</li> | <li>15’’ – still some colonies on UV-part of the plate</li> | ||
<li>20’’ – one colony visible on UV-part of the plate</li> | <li>20’’ – one colony visible on UV-part of the plate</li> | ||
- | <li>30'' – | + | <li>30'' – no growth on UV-part of the plate</li> |
</ul> | </ul> | ||
</ul> | </ul> | ||
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<b>CONCLUSION</b> | <b>CONCLUSION</b> | ||
<br> | <br> | ||
- | With a 30’’ UV exposure, the cells in which <i>recA</i> is successfully deleted will not survive on the | + | With a 30’’ UV exposure, the cells in which <i>recA</i> is successfully deleted will not survive on the UV-treated plate. </p> |
<h1> CIChE </h1> | <h1> CIChE </h1> |
Revision as of 02:26, 4 October 2013
Plasmids containing T7-ccdB![]()
We constructed plasmid pSB6A1-T7ccdB for use in CIChE. Through this plasmid, pressure can be put on the CIChE strains simply by adding IPTG.
Strains with ccdA-gfp constructThe following strain containing the construct to be duplicated (ccdA-gfp) was constructed (KI experiment 1): We also dispose of a backup version of this strain, constructed by Inbio in case our own experiment failed (called MDM.Cm). We included this second version in our further experiments as to test possible differences between the two versions. CIChE Strains
The plasmids and strains mentioned above were used in transformations (experiment 3) to obtain strains containing all elements necessary to perform chromosomal evolution. The following strains were constructed:
UV Test Phage transduction was used to perform the deletion of the recA gene after chromosomal evolution. To check whether the gene was successfully knocked out, an UV test was developed. The UV light puts a certain amount of stress on the bacterial cells. In respons to this stress, an SOS pathway is triggered in which recA plays an important role. Cells in which the recA gene is deleted will not survive on the UV-treated plate. Colonies in which recA was successfully deleted still can be found on a non-treated backup plate.
CIChE
Recommendations for further research
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