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Team:Potsdam Bioware/Biobricks/All Biobricks
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| - | + | <tr><td>[http://partsregistry.org/Part:BBa_K929002 BBa_K929002] modified_AID with CMV and hGH-polyA</td> | |
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For strong expression of the previously described part an CMV-promoter and hGH polyadenylation sequence were added. | For strong expression of the previously described part an CMV-promoter and hGH polyadenylation sequence were added. | ||
| - | + | </td> | |
| + | </tr> | ||
| + | <tr><td>[http://partsregistry.org/Part:BBa_K929003 BBa_K929003]modified AID with CMV, hGH-polyA and eGFP</td> | ||
| + | <td> | ||
The modified AID was fused with eGFP to check 1.)the tranfection success 2.)the cellular lokalization of the improved AID and 3.) to select transfected cells via FACS. | The modified AID was fused with eGFP to check 1.)the tranfection success 2.)the cellular lokalization of the improved AID and 3.) to select transfected cells via FACS. | ||
| - | + | </td> | |
| + | </tr> | ||
| + | <tr><td>[http://partsregistry.org/Part:BBa_K929004 BBa_K929004] modified_AID+eGFP</td> | ||
| + | <td> | ||
For strong expression of the previously described part an CMV-promoter and hGH polyadenylation sequence were added. | For strong expression of the previously described part an CMV-promoter and hGH polyadenylation sequence were added. | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
====Characterization:==== | ====Characterization:==== | ||
Revision as of 20:05, 26 September 2012
Contents |
All Biobricks
As the project is divided into three subproject, the resulting biobricks are listet in the three categories. Furthermore the Biobricks for the new RFC "Potsdam Standard" are listed.
Antibody Biobricks
Mutation Biobricks
All the Mutation Biobricks are improved/modified versions of the wildtype AID that was available as a Biobrick (BBa_K103001). AID is known to be responsible for somatic hypermutation and the class-switch recombination of immunoglobulin in B cells. This enzyme of 28 kDa originally occurs in B cells but does also show activity after transfection into CHO cells. AID induces the deamination of cytidine to uridine at actively transcribed single strand DNA. The replacement of cytidine by uridine leads to a mismatch during DNA replication and integrates a single base substitution predominantly in the immunoglobulin genes.
Therefore this enzymes was used in our project for diversification of antibodies.With PCR and cloning we developed AID`s for the differnt requirements.
| BBa_K929000 AID with CMV promoter and hGH-polyadenylation signal sequence |
CMV-promoter & hGH polyadenylationsequence were added to the wildtype AID for strong expression. |
| BBa_K929001 modified_AID without NES, with NLS and Kozak sequence |
To improve the mutation rate the AID was located into the nucleus, because there it mutates transcribed DNA. Therefor an aditional NLS(Nuclear Localization Sequence)was added and the naturally occurring NES(Nuclear Exprot Sequence) was removed.For stronger Expression an Kozak Sequence was added. To fuse modified AID((C-terminal of mod. AID) with RFC 25 parts, we added an AgeI restriction site. |
| BBa_K929002 modified_AID with CMV and hGH-polyA |
For strong expression of the previously described part an CMV-promoter and hGH polyadenylation sequence were added. |
| BBa_K929003modified AID with CMV, hGH-polyA and eGFP |
The modified AID was fused with eGFP to check 1.)the tranfection success 2.)the cellular lokalization of the improved AID and 3.) to select transfected cells via FACS. |
| BBa_K929004 modified_AID+eGFP |
For strong expression of the previously described part an CMV-promoter and hGH polyadenylation sequence were added. |
Characterization:
Virus Biobricks
Potsdam Standard Biobricks
